text:
Biography of Edward C. Gibbs
North Central Ohio Biographies

Edward Clark Gibbs. One of Medina's most representative young business men is Edward Clark Gibbs, who is identified with the Gibbs Motor Company, local distributors of Chevrolet automobiles. He is also a veteran of the World War. Mr. Gibbs was born at Brunswick, Ohio, June 15, 1893, the son of Farnum Henry and Ellen L. (Blakeslee) Gibbs.

Farnum Henry Gibbs was born on a farm in Cuyahoga County, Ohio, Sept. 12, 1858. He was the son of Farnum M. Gibbs, who was widely known throughout this section of the country as a minister of the Christian Church. He was born about 1834. As an evangelist, Reverend Gibbs was popular and his dominant personality exerted its influence for more than 40 years. He was assisted in his religious work by his daughter, Lillian, who accompanied him on his numerous tours. It is said that Harold Bell Wright became a member of the faith of Reverend Gibbs and later attended Hiram College, where his talent as a novelist was developed. Reverend Gibbs died in 1908 and is buried in Brunswick Cemetery.

Farnum Henry Gibbs, father of the subject of this sketch, became a farmer and merchant, and for many years conducted a store at Brunswick. He married Miss Ellen L. Blakeslee, a teacher at Weymouth, Medina County. She was the daughter of Ashur and Betsy Louise (Perkins) Blakeslee. Her parents died when she was a very small child and she was reared by her maternal grandparents, Mr. and Mrs. Edward Perkins. By a former marriage, Farnum H. Gibbs had a daughter, Nellie, who is the wife of G. R. Moxley, of Brunswick. To Farnum H. and Ellen M. (Blakeslee) Gibbs the following children were born: Eldredge F., who is assistant cashier of the Old Phoenix National Bank, Medina; Calista A., married Harry E. Vaughn, lives near Brunswick; Anna L., married Harry Lincoln, lives at Medina, and he is leader of the Medina Symphony and Medina Band, and well known as a maker of violins and cellos; Edward Clark, the subject of this sketch; Earl B., a sketch of whom appears elsewhere in this history; Esther, married Russell Kinton, lives at Cleveland; and Ellen, attends Hiram College.

Edward Clark Gibbs received his education in the public schools of Brunswick and Medina, from which he was graduated in 1912. He received the degree of Bachelor of Arts at Baldwin Wallace College in 1917, and throughout his college career had devoted his time also to the sale of automobiles, thus defraying his college expenses and accumulating the capital from which his present business was financed without any assistance from other sources. At the outbreak of the World War he enlisted for service in the Motor Transport Corps and was sent to Roys Automobile School, Kansas City, Mo., as an instructor, and in charge of teaching enlisted men to drive trucks. He was later transferred to the Signal Corps and sent to France, where he served throughout the remainder of the war period as master signal electrician. He was discharged in March, 1919, and immediately engaged in the automobile business at hjs present location, South Court and East Smith Road. Mr. Gibbs has been associated in business with his brother, Earl B. Gibbs, since 1921, and they are recognized as progressive young business men of the city. During his career as local dealer for the Chevrolet cars, Mr. Gibbs has completed the sale of 2,818 new automobiles and 5,226 used ears.

On June 22, 1920, Mr. Gibbs was united in marriage with Miss Mabel M. Deuble, the daughter of George and Emma Deuble, of Brunswick. They are the parents of two daughters, Miriam and Barbara.

Mr. Gibbs is a Republican, a member of the Church of Christ, and is affiliated with Medina Lodge, F. & A. M., No. 58; Medina Chapter, R. A. M., No. 30; Wooster Council, No. 13; Independent Order of Odd Fellows; American Legion; and Kiwanis Club. As chairman of the committee on underprivileged children for the Kiwanis Club, Mr. Gibbs was active in the establishment of the first open air camp for children, which has now become an institution with the club. He always takes an active part in the Medina Community Chest Campaign and was chairman of the 1929-30 campaign.

Mr. Gibbs is an ardent sportsman, and especially interested in hunting and fishing. He takes numerous trips into Canada and the Rocky Mountain region, and has in his possession countless interesting trophies brought from these expeditions. He also has a number of stereopticon films which he uses to illustrate lectures which he has been asked to deliver upon a number of occasions.

The Gibbs family lives at 504 Wadsworth Road, Medina.

Also see Earle B. Gibbs. (His brother)

History of North Central Ohio
Embracing Richland, Ashland, Wayne,
Medina, Lorin, Huron and Knox Counties
BY: William A. Duff
Historical Publishing Company
Topeka-Indianapolis 1931

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North Central Ohio Biographies

Names A to C
Names D to G
Names H to K
Names L to P
Names Q to S
Names T to Z

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For all your genealogy needs visit Linkpendium

Family Tree Maker 2012


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text:
 Tic tac toe in Android free project download|online free project download

Android projects

This category consists of android projects list, android projects with source code which can be downloaded by final year students. Here we listed out collection of android projects.

Tic tac toe android project source code

Sudoku game is one the popular game played by most number of users all over the world. Because of it puzzle words it has created interested in users. Initially this game use to be available only for systems.

But in present trend mobile market had rapidly increased in developing mobile games. There are many mobiles operating system providers like android, windows, bada. In present trend android is the leading operating system for mobiles. This is because of free apps and games provided in app store for free download.

Tic tac toe android project source code and report download.

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Men's Felix 6" WP Duck Boot
Men's Felix 6" WP Duck Boot
Color: Black Multi Leather
  • $189.95
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when it's back.

Product & Brand Information

With the classic duck-boot look crafted completely in full-grain leather, the Wolverine Felix Waterproof Duck Boot provides the adventurous outdoor style in a slightly more refined realization. Adventure ready capability is maintained with a waterproof interior membrane and a pliant rubber outsole for dry feet and reliable traction in wet conditions.
  • Item # 539629
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What is Bone Marrow Cancer?

By  ,  Onlymyhealth editorial team
May 18, 2012
4.8 / 5(4 Ratings)

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The soft tissue in your bones is known as bone marrow. Stem cells form soft tissues which form the bone marrow. The bone marrow cancer is the result of the stem cells becoming cancerous. Although bone marrow cancer is not contagious, its exact cause is not known. Doctors cannot explain why some people get affected by bone marrow cancer while others do not.

That is why the prevention of bone marrow cancer is not easy. Knowledge of exact cause would allow for the treatment to be developed, but since that is not the case, prevention is not possible. Certain diseases inherited from the parents are responsible for the bone marrow cancer. The risk factors of this form of cancer are unavoidable as they are genetic.

Some of the causes of bone marrow cancer are:

  • Chronic condition of the bone.
  • Family history of bone marrow cancer.
  • Umbillical hernia at the time the child was born.
  • The DNA of stem cells develops some sort of defect.
  • If it is the case of child below five years of age, effect of retinoblastoma or cancer in the eye can be bone marrow cancer.
  • People in old age are more likely to develop degenerative disorders such as this type of cancer.
  • Some genetic disorders which are quite rare such as Li-Fraumeni syndrome.

You cannot prevent most cases of bone marrow cancer but some precautionary measures would still reduce the risk of it occurring. You need to be vigilant about the occurrence of this form of cancer. The symptoms of this form of cancer are excruciating pain in the bones and fatigue.

Hard lumps and swelling along with stiff joints along are some of the common symptoms of bone marrow cancer. Fractures, reduced immunity, anaemia and minor injuries are also the symptoms of bone marrow cancer.

The patient should be always aware that the symptoms can vary from one to another. You will need to visit the doctor to rule out or diagnose the presence of cancerous cells in your body. Malignancy or the presence of cancerous growth in the body is indicated by weight loss, loss of appetite and a general weakness. You need to stay in touch with your doctor for the recommendations and follow them strictly.

In most of the cases, bone marrow cancer is caused by an overgrowth in a particular organ. If point of origin of the disease is found out, treatment can be done promptly.

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Liverpool sign Sporting Lisbon defender

Liverpool have confirmed the signing of defender Tiago Ilori from Portuguese side Sporting Lisbon.

The 20-year-old was born in London but has represented Portugal at youth level.

Ilori told Liverpool’s official website: “It feels great being a Liverpool player – coming to the Premier League has always been a dream for me.

“I’m going to try and achieve everything I can. I want to play as much as possible and we’ll see what the future holds. I will give my all and try my best to help the team.”

“I know the history of the club and the Premier League is the best league in the world, in my opinion,” continued Ilori. “Liverpool, have a great team and I think it’s the place for me to be at the moment.

“I know there have been some great teams here in the past and the club has won a lot of trophies and I want to be a part of it.”

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Summer 2016 is set to be a very exciting time for football fans, as this is when the long awaited Euro 2016 tournament kicks off.

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text: ospfd, bgpd and the routing table instead, we want them to install the routes they learn blindly use RTF_MPATH, so there can be more than one route per prefix store the route origin (BGP/OSPF/...) with it kernel knows that static takes precedence over bgp, etc same for other routing daemons, if anyone wants to hack on IS-IS, drop me a mail :) extensive changes, not as easy as it sounds. hope to attack that soon.


text:
We are a Spanish company leader in providing IT consulting and global solutions based on Linux platform and Open Source software. We also assume low level C/C++ development projects, Linux Kernel optimizations, and demands of customization processes applied on Open Source applications.

We offer to our customers and partners our value-added services and highly qualified personnel, so we are involved in many critical projects with some of the biggest companies and public agencies in Spain.

Country: Spain

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  • Alkacon OCEE 5.5.0 for OpenCms 10.5.0 available

    Dec 9, 2016 - OCEE 5.5.0 supports OpenCms 10.5.0 and brings seveal improvements to the LDAP Connector. It also adds more shell commands for remote cluster administration, and fixes some issues. Updated OCEE packages for 10.0, 9.x and 8.x based OpenCms installations are also available.

  • Introducing OpenCms 10.5

    Nov 30, 2016 - OpenCms 10.5 focuses on site localization, adding tools for assisted translation of pages and sitemaps. Many usability improvements have been made for the Page Editor and the Explorer. OpenCms 10.5 also ships a much improved version of the Apollo Template. Based on Bootstrap, the Apollo Template is fully customizable without coding.


text:
‘Tis the very nature of parenthood to view one’s children as exceptional.

Another aspect of the condition is spending time in the company of other parents, some of whom have yet to master the art of self-restraint. Their babies are the most physically adroit, their toddlers the most generous, their elementary schoolers the most culinarily daring.

Pride in one’s children’s gifts is understandable. A straightforward brag or two is permissible. But after that, I’d really like some corroborating evidence, such as the Honourable Daines Barrington’s account of meeting a “very remarkable young musician” whose father had been dragging him around the continent on a 3-and-a-half year concert tour.

Clearly, Johannes Chrysostomus Wolfgangus Theophilus Mozart (1756-1791) was a very accomplished kid, but the term “prodigy” must have stuck in Friend of the Royal Society Barrington’s craw, even after he’d attended some of the boy’s public performances. Determined to let science be the judge, he devised a series of on-the-spot challenges designed to evaluate the boy’s musicianship beyond the rigorous practice schedule imposed by his disciplinarian father. (We all saw Amadeus, right?)

Barrington’s detailed description of these experiments would make a gimcrack Science Fair project for any little Einstein smart enough to get through 18th-century typography without throwing a tantrum because the s’s all look like f’s (see the 18th century text below). It might take a recreational mathemusician on the order of Vi Hart to truly appreciate the complexity of the tasks that Barrington assigned his young subject (something to do with having him play five contradictory lines simultaneously…).

I can interpret the data with regard to some of Barrington’s other findings. Mozart, he wrote, not only looked like a young child, he ditched his harpsichord to chase around a cat and ran about the room “with a stick between his legs by way of a horse.” So it’s not the classical period equivalent of a stage mom lying about her kid’s age to better his chances at an audition. Actually, it’s always a relief to hear about these super-kids acting like… well, my kids.

Read Barrington’s letter to the Royal Society — Account of a Very Remarkable Young Musician — in its entirety here or below. And by all means share it with the worst offenders on your PTA.

via Rebecca Onion and her Slate blog, The Vault

Related Content:

Newly Discovered Piece by Mozart Performed on His Own Fortepiano

Great Violinists Playing as Kids: Itzhak Perlman, Anne-Sophie Mutter, & More

The Musical Mind of Albert Einstein: Great Physicist, Amateur Violinist and Devotee of Mozart

Ayun Halliday provided an honest account of homeschooling her 12-year-old son in The East Village Inky #51 Follow her @AyunHalliday

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text:
Re: Remove an objectclasse during syncrepl

> Hi all,
> We have a brand new ldap server that we are going to replicate with an
> outside replica, for an extranet purpose.
> During this replication, we would like to remove the "posixaccount"
> objectclass to only let the "inetorgperson" and "top" ones so we didn't
> need to put passwords or anything not needed
> I heard about slapo-rwm, but it seems to be buggy

Let "buggy" aside (it's not "buggy" but it may have interaction problems
with slapo-syncprov(5)).  Or, if you notice a bug, please submit an ITS.

> I'm sure that some of you all ready have done that, maybe there is a
> better way.

You should use the primary mechanism syncrepl provides for this purpose:
the filter, the attribute list and ACLs.  You can simply hide attributes
related to posixAccount in a specific set of ACLs that are only triggered
by the replicator's identity.  Something like

access to attrs=objectClass val=posixAccount
	by dn=cn=replicator none
        by * break

access to attrs=@posixAccount
	by dn=cn=replicator none
        by * break

Note: you may need to craft that a little bit if posixAccount also
contains stuff used by other objectClasses you don't want to be filtered


text:
Yazoo-Mississippi Delta Levee District

Agencies 2010

AgencyNo. of Reports Listing Agency*
US Senate4
US House of Representatives4
Army Corps of Engineers4
Federal Emergency Management Agency3
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*Each quarterly filing is treated as a separate report.

Feel free to distribute or cite this material, but please credit the Center for Responsive Politics. For permission to reprint for commercial uses, such as textbooks, contact the Center: info@crp.org

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Issues, 2008

Issue Specific IssuesNo. of Reports*
Fed Budget & Appropriations88
Energy & Nuclear Power66
Science & Technology45
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*Each client may report lobbying on multiple issues.

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text:
Capacity building for local development, a comparative study of formal and informal organizations in Gondar, Northern Ethiopia

by Mequanent, Getachew

Abstract (Summary)
This thesis explores the concephial and methodological questions of local capacity building. The issue of capacity building has to do with enhancing the role of local organizationsin the development process. It has ken argued that strong organizations would create the social and material basis of support for development initiatives, including the structuresdemanded by decentralized power, institutions, and resources. But,too ofien there has been little done to assess the range of organizational capabilities which are the determinhg factors for the effectiveness and eficiency of their performance functions. On the basis of the case study of " formal " and "informal" organizations in Gondar, Northem Ethiopia, this thesis argues that local organizations are created and sustained by dynamic processes of socio-economic, cultural and political interactions, which, in tum, require individuals to engage in a systematic mode of interaction to b e and rehe relationships arnong themselves and with outsider agencies. The case study examines the roles, structures and fûnctions of these organizations to undestand how they actuaily work and also how the environment that is external and intemal to organizations can increase or limit the range of their capabilities as social agencies of change in local society. The study also develops a framework to assess the importance of existing and potentid organizational capacities to initiate and coordinate development work. In doing so, it attempts to identify both "capacity utilization " problems and " capacity building " needs. The problem of utilizing existing capacities emanates frorn the topdown, technocratie practices which rarely take account the vital role of local organizations in the development process. The capacity building issue draws attention to the need for developing local knowledge-base and technical skills that match people's expectations in the development process. The study concludes by proposing a regional planning approach that helps to define areas of responsibility for local organizations, so as to complement their des with programme and project initiatives.
Bibliographical Information:

School Location:

Source Type:Master's Thesis

Date of Publication:01/01/1998

© 2009 All Rights Reserved.


text:
BISC219/F13: RNAi Lab 8

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m (Lab 8: Series 3- Creating the feeding strain of bacteria for RNAi in C. elegans)
Line 28: Line 28:
In order for RNAi to be successful we must insert the plasmid expressing dsRNA of our ''C. elegans'' gene of interest into the bacteria that will make lots of double stranded RNA.  We will then feed the bacteria that is full of double stranded RNA specific to our gene of interest to our worms.  The process of inserting the plasmid into bacteria is called transformation.  Transformation, either chemically or electrically, opens up pores or channels in the bacterial membrane that allows for uptake of the plasmid DNA.

In order for RNAi to be successful we must insert the plasmid expressing dsRNA of our ''C. elegans'' gene of interest into the bacteria that will make lots of double stranded RNA.  We will then feed the bacteria that is full of double stranded RNA specific to our gene of interest to our worms.  The process of inserting the plasmid into bacteria is called transformation.  Transformation, either chemically or electrically, opens up pores or channels in the bacterial membrane that allows for uptake of the plasmid DNA.

Precise pipetting is required for transformation to be effective.  To ensure we are all pipetting properly and that our pipettes are properly calibrated follow the procedure below.
Precise pipetting is required for transformation to be effective.  Your instructor will demonstrate the correct technique. If you have any questions, please consult with her/him before proceeding with any micropipetting.
== Part 1:Calibration of Micropipettes ==
== Part 1:Calibration of Micropipettes ==

Revision as of 09:53, 6 November 2013

Lab 8: Heat Shock Proteins (HSP's) and Heat Shock Factor -1 (HSF-1)

Background materials for HSP's, HSF-1 and the reporter assay are provided by Cassandra Theusch and Peter Kuhn; Elisabeth A. Cox and Theresa M. Grana.
Although “heat shock proteins” are best characterized in associated with temperature, mounting evidence suggests that heat shock proteins are expressed and play vital roles during many types of cell stress. Heat shock proteins work by helping other proteins fold properly and have been classified in a functional category of proteins called molecular chaperones. Molecular chaperones catalyze the folding of a newly-translated peptide into the secondary and tertiary structures that characterize the mature protein product. Improperly-folded proteins are unstable, prone to aggregation, and dysfunctional.

Correct protein folding is extremely important for protein and cell function. . Alterations in protein structure, including secondary and tertiary structure, lead to altered protein function. Similarly, improperly or incompletely folded proteins are likely to be dysfunctional and lead to complications for the cell. It should come as no surprise that improper protein folding in cells of the human body can lead to severe diseases. For example, transmissible spongiform encephalopathies (TSE’s), such as mad cow disease, chronic wasting disease, and Creutzfield-Jakob disease, are the result of improperly-folded proteins called prions. A buildup of prions interferes with cell functions and leads to apoptosis (programmed cell death). Other protein folding diseases exhibit similar pathological mechanisms. Alzheimer’s disease is thought to result from the aggregation of improperly-folded amyloid protein. Amyloid plaques accumulate in the brain and damage surrounding neurons as they grow. Certain types of cancer are associated with molecular chaperone dysfunction, which allows tumor cells to grow abnormally and survive adverse conditions.

Heat shock factor 1 (HSF-1) is a transcription factor which is required for transcription/expression of genes encoding many subtypes of heat shock proteins. Under normal cell conditions, HSF-1 expressed in the cytoplasm migrates occasionally, in small amounts, to the nucleus to activate transcription of genes encoding heat shock proteins to carry out normal cell functions. During a cell’s response to stressful conditions, HSF-1 is activated in large amounts, polymerizes, and migrates from the cytoplasm to the nucleus to search out recognized promoter sequences. This high level expression of heat shock proteins allows for refolding of proteins following stress and recovery of protein function. The specific mechanisms and molecular triggers of HSF-1 activation and migration are still unclear. Some evidence points to certain subtypes of heat shock proteins binding with and phosphorylating HSF-1 during the stress response. Yet other HSP’s may have roles inhibiting HSF-1, although scientists are still working to identify all the facets of HSF-1 regulation. More broadly, the types of stressful cellular stimuli which activate HSF-1 also have yet to be elucidated. This is a hot area of research currently, where scientists are hoping to answer some important questions about how cells respond to stressful stimuli. Perhaps your investigations will contribute to this research!

GFP and the reporter assay

A reporter assay is commonly used to assess the activity of transcription factors within a cell. You will use reporter assays to determine functional expression of the transcription factor HSF-1. A reporter assay exploits the expression of a measurable (or visible) protein, in this case GFP. The gene encoding GFP is transgenically engineered to lie downstream of a promoter to which the transcription factor binds and initiates transcription. This engineered “transgene” can be incorporated into the genome of an organism, such as C. elegans. Therefore, when conditions are right and the transcription factor’s expression is high enough, reporter gene is transcribed, and this can be measured by the amount of reporter protein produced.

Green fluorescent protein (GFP) is a commonly-used reporter protein, and, as its name suggests, is a fluorescent protein used in countless ways to visualize and quantify cellular molecules and processes. Originally, GFP was identified in and isolated from the marine jellyfish Aequorea victoria. However, modern advances in molecular biology have produced transgenic forms of GFP to more aptly suit certain experimental requirements. Due to the explosion of discoveries through experiments using GFP, the 2008 Nobel Prize in Chemistry was awarded to Martin Chalfie, Osamu Shimomura, and Roger Tsien for the initial discovery of GFP.

Your experiments will use a transgenic C. elegans strain which has been engineered to contain within its genome a reporter gene. This reporter construct encodes GFP under the control of a hsp-16.2 promoter. (Normally, expression of hsp-16.2 genes results in HSP-16.2 protein production.) HSP-16.2 is one subtype in the family of heat shock proteins, and therefore, it carries out molecular chaperone duties when it is present in the cell. HSF-1 binds to and initiates transcription via the promoter to hsp-16.2. Genetic engineers merely chopped out the promoter to hsp-16.2, pasted it onto the gene encoding GFP, and inserted the combo into a worm’s genome – a reporter gene was born!

Image:Hsp image.jpg

Figure 1: Construction of the GFP reporter transgene. The promoter region of hsp-16.2 was pasted onto the gene encoding GFP. Therefore, transcriptional activity at the hsp promoter can be measured by the levels of GFP in the cell.

Under normal conditions, HSF-1 is present in cells, but its low level of activation cannot be directly measured using the techniques you will use in lab. When a cell is stressed, HSF-1 is phosphorylated and activated in the large amounts measurable using this GFP reporter assay. Activated HSF-1 up-regulates the transcription of the hsp16.2/gfp transgene, which allows measurement of activation through detection of GFP using fluorescence microscopy.

Lab 8: Series 3- Creating the feeding strain of bacteria for RNAi in C. elegans

In order for RNAi to be successful we must insert the plasmid expressing dsRNA of our C. elegans gene of interest into the bacteria that will make lots of double stranded RNA. We will then feed the bacteria that is full of double stranded RNA specific to our gene of interest to our worms. The process of inserting the plasmid into bacteria is called transformation. Transformation, either chemically or electrically, opens up pores or channels in the bacterial membrane that allows for uptake of the plasmid DNA.

Precise pipetting is required for transformation to be effective. Your instructor will demonstrate the correct technique. If you have any questions, please consult with her/him before proceeding with any micropipetting.

Part 1:Calibration of Micropipettes

Before we begin our molecular genetic experiments, we will practice (AGAIN) properly handling and using the micropipettes.

  1. To calibrate your P1000, P200, and P20 micropipets, label 6 microfuge tubes (1-6) and weigh them. Record the weights in the table below.
  2. Following the table below, pipet the specified volumes into the pre-weighed microfuge tubes prepared above and then re-weigh them. Record all weights.
  3. Calculate the weight of the water in grams. 1000 microliters of water should weigh 1 gram at room temperature.
  4. If the water in any tube weighs more or less than 1 gram, ask your instructor for help. If your calibration is significantly off after several repeated attempts, your pipet (or your technique!) may need adjustment.

Tube # Tube Pre-Weight Vol. in μL using P20 Vol. in μL using P200 Vol. in μL using P1000 Weight of Tube + Water in grams Weight of Water in grams
200 (5 times)
20 (5 times)

For a Word™ format protocol: Media:Protocol for Micropipet Calibration.doc

To recap what has been done in order for your to be ready to do today's work:
Yesterday you inoculated a few milliliters of LB broth containing E. coli that we are sure have maintained our genetically engineered plasmid, pPD129.36, modified to contain an antibiotic resistance gene to ampicillin and all or part of the C. elegans hsf-1 gene that you want to investigate. You added ampicillin to the broth to ensure that the plasmid DNA would be maintained by the cells. Overnight the bacteria have grown to high density and the plasmid DNA has undergone many replications. However since you started with a single colony of bacteria and that colony grew from a single transformed cell, all the copies of the plasmid DNA in your overnight culture should be identical (“clones” of one another). To isolate the plasmid DNA from this bacterial strain that expresses C. elegans hsf-1 , you will perform what is commonly called a “mini-prep”. This term distinguishes the procedure from a “maxi-” or “large scale-prep” which involves a larger volume of cells and additional steps of purification. The overall goal of each “prep” is the same--to separate the plasmid DNA from the chromosomal DNA so that a certain gene on the plasmid DNA can be studied further.

Synopsis of Today's Work
Today you will transform the isolated plasmid DNA, pPD129.36+hsf-1, into an IPTG inducible strain of E. coli called HT115(DE3), and spread the transformed bacteria onto LB agar media supplemented with ampicillin (resistance conferred by a gene expressed from the plasmid).

Part 2: Plasmid DNA Isolation

What's Happening in Your MiniPrep
To isolate plasmid DNA from an overnight culture of cells, the media is removed from the cells by centrifugation. The cells are resuspended in “Solution I” which contains Tris to buffer the cells and EDTA to bind divalent cations in the lipid bilayer, thereby weakening the cell envelope. Upon the addition of “Solution II,” the chromosomal DNA and the plasmid DNA are denatured by the sodium hydroxide, and the cellular proteins and lipids are dissolved by the detergent, sodium dodecyl sulfate (SDS). The pH of the solution is returned to neutral by the potassium acetate in “Solution III.” At neutral pH the SDS precipitates from solution, carrying with it the dissolved proteins and lipids. In addition, the DNA strands renature at neutral pH. The chromosomal DNA, which is much longer than the plasmid DNA, renatures as a tangle that gets trapped in the SDS precipitate. The plasmid DNA renatures normally and stays in a water based solution. In this way the plasmid DNA is separated from the chromosomal DNA, the proteins and the lipids of the cell. Plasmid DNA can be precipitated out of solution in absolute ethanol and then put back into solution (in an appropriate concentration) in water. The ingredients and concentrations of a stock solution of all reagents (such as Solutions I, II, and III) can be found in the Media Recipes section of this section of the wiki.

Plasmid DNA Isolation Only one of your two overnight cultures should appear cloudy with bacterial growth. If your control is cloudy, please inform your instructor. (You can find the ingredients and concentrations for ingredients of Solutions I, II, and III in the Series 3 Media Recipes page.

  1. To start the plasmid DNA isolation, label 2 microfuge tubes with pPD129.36+hsf-1 (our plasmid name + our C. elegans gene), plus your initials and team color.
  2. Check to be sure that your cells have not settled. If they have, mix the cultures but do NOT invert them because they will leak. Then pour some of the overnight culture so that both microfuge tubes are almost full. If you are nervous about pouring the bacteria, you can pipet 750 microliters into each tube twice, so that there is a total of 1.5 ml in each tube. The exact volume doesn’t matter but the tubes should be quite full when you close the cap.
  3. Place the tubes in the room temperature microfuge so that the hinges of each cap is facing out. Paying attention to this small detail will help you know where in the tube to find your pellets. While it is not essential to do this in this step, it is a good habit to get into since sometimes pellets are very small and hard to see. Be sure your tubes are balanced, then spin the tubes for two minutes at 8,000 rpm. Check the rcf speed of the centrifuge while it is spinning (by hitting the toggle that changes the readout) and record the rcf’s in your lab notebook. Relative centrifugal force (rcf) is what should be used in M&M to describe your speed since it is universal and rpm is rotor and centrifuge dependent.
  4. If the supernatant is not clear, recentrifuge until it is. When the supernatant is clear, pour it into the waste beaker that is on your bench, then flick the tube with the cap open to remove the last few drops of liquid. The cell pellet will not fall out.
  5. Resuspend your cell pellets COMPLETELY in 100 microliters of solution I. Pipet up and down until ALL of the cells are uniformly suspended in Solution I. Be sure to change tips between samples. Leave the cells at room temperature as you prepare solution II.
  6. To make solution II, mix 500 microliters of 2% SDS with 500 microliters of 0.4M NaOH in a microfuge tube. Close the cap and invert the tube several times to mix the contents. Add 200 microliters of solution II to each miniprep and invert the tubes five or six times to mix. In some cases the minipreps may appear to "clear" but don't worry if you don't see a big change in yours. Place the tubes on ice for five minutes.
  7. Add 150 microliters of solution III to each miniprep and immediately vortex each tube for 10 seconds with your vortex set at the highest setting. White clumps should appear in the solution after you vortex it. Place the tubes in the room temperature microfuge and spin for 3 minutes at the HIGHEST speed.
  8. While the minipreps are spinning, label another set of microfuge tubes with the name of the plasmid (pL4440+C. elegans gene name)and your initials or team color. The new tubes can be left at room temperature.
  9. A white pellet should be visible when you remove your minipreps from the microfuge. Use your P1000 to transfer 400 microliters of each supernatant to the appropriate clean microfuge tube. It's OK to leave some of the supernatant behind. Try to avoid transferring the white pellet.
  10. Add 1 ml of room temperature 100% ethanol to each new tube. The tubes will be quite full. Close the caps and invert the tubes at least five times to thoroughly mix the contents.
  11. Spin the tubes in the room temperature microfuge for 2 minutes at HIGHEST speed. It is important to orient your tubes so that the hinges are up this time, as the pellets are expected to be barely visible.
  12. Pour off the supernatants into the waste beaker on your bench then flick out the last few drops. The tubes do not have to be completely dry.
  13. Carefully, wash the pellets with 500 microliters of 80% ethanol. (You may need to make a few mls of 80% ETOH from the 100% ethanol stock provided). When you wash the pellets, add the 80% ethanol slowly and carefully, so that the liquid flows down the side of the tube away from the pellet. After the ethanol has been dispensed, immediately remove it with the same tip, making sure to keep the tip on the side of the tube that doesn't have your pellet. Some liquid will remain in the tube, and it can and must be removed using your P200, set to 100 microliters. Try to remove as much of the ethanol as you can (without removing your pellet!). The pelleted DNA will have to be completely dry before you add water or your DNA will not go into solution.
  14. After all the DNA pellets have been washed, put your tubes with their caps open in the SpeedVac that is at the back of the lab. If you were able to remove most of the ethanol with the P200, then after two or three minutes the last drops of ethanol will have evaporated and the pellet will be barely visible. If the DNA pellet is invisible please see your instructor to determine how much water to use in the next step.
  15. If you have a visible pellet of DNA, add 30 microliters of sterile water to each tube and vortex each tube enough for all of the DNA to be off the wall of the tube and into solution. The pelleted plasmid DNA must dissolve completely in the water. The liquid can be brought back to the bottom of the tubes by spinning for a few seconds in the microfuge. You can then combine the dissolved plasmid DNA into one of the two tubes so that you have approximately 60 microliters of plasmid DNA as one aliquot. Store the DNA on ice until you are ready to use part of it and then give the rest to your instructor to freeze. DO NOT DISCARD ANY OF YOUR PLASMID ISOLATE!

Part 3: Measuring the concentration of plasmid DNA using the NanoDropper

Proper transformation requires the concentration of bacteria to plasmid DNA to be right. We will use a nano-dropper to determine the concentration of the plasmid DNA from a previous plasmid isolation procedure.

Use the ThermoScientific NanoDrop Spectrophotometer in L308 to measure DNA by taking Absorbance at A260nm. This spectrophotometer uses only 1 microliter of sample and does not require cuvettes. The sample is held in place by fiber optic technology and surface tension that holds the sample in place between two optical surfaces that define the pathlength vertically and dynamically. Measurement can be assessed in a range of 2 to 3700nm/microliter dsDNA. These are expensive machines so make sure you follow the directions carefully and ask your instructor for guidance as needed.

More information is available from the manufacturer's website at: |

Using the Nanodropper
1. Clean the upper and lower optical surfaces of the sample retension device by pipetting 2 microliters of clean deionized water onto the lower optical surface. Close the lever arm and tap it a few times to bathe the upper optical surface. Lift the lever arm and wipe off both optical surfaces with a Kimwipe.

2. Open the NanoDrop software from the Desktop of the computer and select the nucleic acids module.

3. Initialize the machine by placing 1 microliter of clean deionized water onto the lower optic surface, lower the lever arm, and select initialize from the NanoDrop software. Once initialization is complete (~10sec.), clean both optical surfaces with a Kimwipe.

4. Perform a blank measurement by loading 1 microliter of purified deionized water.
Note that as in traditional spectroscopy, the blank will be subtracted from subsequent measurements. If you want to determine the contribution of a specific buffer or diluent, measure the buffer first using distilled water as a blank. If the buffer does not contribute to the A 260nm reading, then deionized water is fine to use as the blank. The water or buffer should always be measured to be sure that the instrument has been zeroed properly. The measurement of water or buffer should be zero or very close. All measurements are automatically normalized to 340nm.

5. Measure the nucleic acid sample by loading 1microliter of sample and selecting "measure". Record your DNA concentration in your lab notebook and on the tube of plamid DNA. Once the measurement is complete. Clean both optical surfaces with a Kimwipe and the machine is ready for the next sample.
You should ensure that the appropriate constant (50 for dsDNA or 40 for RNA) has been chosen. The software automatically calculates the nucleic acid concentration. If the calculation is done by hand, the A260nm is represented as a 1cm path for convenience, even though 1-nm and 0.2nm paths are actually used during the measurement cycle.

Clean Up When the last sample was been measured, clean the sampling device by repeating step 1.

Part 4: Transformation of plasmid DNA into E. coli strain HT115(DE3)

The competent HT115(DE3) bacterial cells are on the instructor’s bench. You will transform some of your plasmid DNA into these bacteria. The cells are very fragile, so treat them gently.

The genotype of the HT115(DE3) cells is: F-, mcrA, mcrB, IN(rrnD-rrnE)1, rnc14::Tn10(DE3 lysogen: lavUV5 promoter -T7 polymerase) (IPTG-inducible T7 polymerase) (RNAse III minus). This strain grows on LB or 2XYT plates. This strain is tetracycline resistant. The most important things about this strain is:

  1. It has an IPTG inducible T7 polymerase. The chemical IPTG (Isopropyl β-D-1-thiogalactopyranoside) is a lactose mimic and is commonly used to induce the production of large amounts of protein. In this case when the cells are incubated with IPTG lots of the polymerase T7 is made. This T7 polymerase binds to T7 specific promoters on the plasmid and transcribes lots of our gene of interest.
  2. RNAse III minus. The strain is lacking the RNAse III enzyme which typically breaks down double stranded RNAs. We want double stranded RNAs to accumulate.

The prep staff prepared these competent HT115(DE3) cells for you using the Inoue Method Media:Inoue bacterial transformation.doc. Reference: Inoue H., Nojima H., and Okayama H. 1990. High efficiency transformation of Escherichia coli with plasmids. Gene 96: 23-28. The cells were made competent to take up free plasmid DNA by this treatment. This treatment makes the cell wall and membrane more permeable and our transformation efficiency much greater but it weakens the cells; therefore you must keep them cold and mix them gently throughout the transformation (no vortexing!).

  1. Obtain a tube containing 50 μL of competent cells from your instructor.
  2. Label the top or the side of the tube with HT115(DE3), pPD12936+hsf-1, and your initials or team color.
  3. Consult with your instructor to determine how much of your plasmid DNA you should add to the cells to start the transformation. When you have determined the correct vol., pipet between 1μL and 10μL of your plasmid DNA to the tube. (The volume you should use is dependent on the conc. you achieved in your mini-prep.) Pipet up and down once to mix the DNA and the cells. Close the cap and let the transformation mixture sit on ice for 10 minutes.
  4. Heat shock by incubating the transformation mix at 42°C for 90 seconds, exactly. This step must be timed exactly. Remove the tube at the end of 90 seconds to your ice bucket while you get your LB ready.
  5. Consult your instructor to determine the volume of warm LB broth you should add (usually between 250-500 microliters) to the transformation mix. LB aliquots should be found in the 37°C mixer/incubator on the green/blue team bench. When pipetting the media, remember to release your thumb on your micropipet slowly, to avoid splashing the liquid on the end of the barrel. The barrel is not sterile and if you see the liquid touch it, then discard the media in the waste beaker and try again with a new tip.
  6. Once you have added the LB, close the cap and invert the tube once or twice to mix the contents. Incubate at 37°C for 30-45 minutes in the bench top incubator/ mixer with occasional gentle mixing.
  7. While the plasmid DNA is being taken up by the competent cells and the new genes provided by the plasmid are being expressed by the bacteria, label one LB + amp agar plate and one LB + amp + tet plate. Label the bottom of the plated with the strain's identity (HT115(DE3)), the plasmid used, the date, your initials and team color. You must label the bottom of the plated since the tops are easily switched. Put these plates in the hood with the blower on and with the lid ajar to dry the surface of the agar for about 10 minutes or until the surface looks dry but is not badly dehydrated.
  8. Once the transformation mix has incubated at 37°C for 30-45 minutes, invert it to mix the contents and pipet 100 microliters of transformed cells onto the center of each of your labeled plates. Pour 5-10 glass beads onto the plates. Put the lid back on and gently swirl the beads all over the agar to spread the transformed bacteria around. When you are done pour the beads into the beaker with disinfectant near the sink.
  9. Replace the lids and leave the agar plates undisturbed for a few minutes to allow for the liquid to be absorbed by the agar.
  10. Once they have dried enough that the surface does not appear wet, invert the plates (lid side down) and incubate them at 37°C for 24 hours. The plates should be incubated with the agar side up so that condensation will not drip onto the surface of the agar and smear the colonies that will be growing there.
  11. Save the remaining transformation mix for 24 hours or until we are sure that there is at least one colony growing on each of your plates.

What would it mean if you had no colonies on your plate? Normally, you would expect to have around 100 pale color colonies on each plate. If you have at least one well isolated colony on the plate, you’re all set. After the 24 hour growth period the plate should be placed in the rack in the refrigerator labeled with your lab day. You will use a single colony from the plate to make an overnight broth culture on the day before next lab. If you have no colonies on one or more of your plates, please notify your instructor right away.

Before leaving lab today, give the rest of your isolated plasmid DNA to your instructor in a labeled microfuge tube. Make sure your tube is labeled with your name, lab day, plasmid name, DNA concentration and color coded with a piece of tape in your team color.

To do on the day before the next lab

You and your partner will return to the lab to make an overnight broth culture of your selected colony as described below. This process will create a sub-culture of many identical copies of the bacteria containing the plasmid carrying the construct to RNAi the gene that you want to study.

  1. Find your plate in the glass front refrigerator in a rack labeled with your lab day. Select a colony to start your overnight culture. At this point, ALL of your colonies should contain your plasmid of interest.
  2. Begin by obtaining two tubes of LB broth (each will have 5 ml of broth) from the refrigerator in the back left hand corner of the room.
  3. Add 5 microliters of the 50mg/ml ampicillin stock (also found in the refrigerator with the broth) to each tube. Calculate the effective concentration of ampicillin that you have in your LB tube (remember V1 x C1= V2 x C2) and record that information in your lab notebook.
  4. Add 5 microliters of the 12.5mg/ml tetracycline stock (also found in the refrigerator with the broth). Calculate the effective concentration of tetracycline that you havein your LB broth tube. Record that info in your lab notebook.
  5. Gently swirl your LB +amp+tet broth to mix the contents.
  6. Label the two sterile glass culture tubes with tape in your team color. Label one with "pPD129.36 hsf-1" and your initials. Label the other with your initials only.
  7. Inoculate the broth with your bacteria by using a sterile disposable loop to scrape your candidate colony off the plate. Be sure not to touch the plate with the loop except on the desired colony and don’t pick up any satellite colonies! Gently swirl the loop in the LB+amp+tet broth - you should be able to see the colony come off the loop. The second tube of broth labeled with just your initials is a control and should not be inoculated with bacteria as it is your control for contamination.
  8. Balance the 2 tubes across from each other on the rotating wheel in the 37C incubator at the front of the room when you come in the door.
  9. Incubate these broth cultures at 37°C overnight. Do not forget to make sure the wheel is rotating when you leave!

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Judging Chaudhry

IT is ungracious to take pot shots at someone walking into the sunset. But to refuse to take stock of the performance of a public office holder as remarkable as Chief Justice Iftikhar Chaudhry and draw lessons for the future is also undesirable.

In measuring up Chaudhry’s tenure as chief justice there are five broad areas that invite comment: judicial independence; suo motu powers; judicial appointments; judicial reform; and Arsalan Iftikhar.

CJ Chaudhry’s lasting contribution to Pakistan is that no one dare mistake the judiciary for being an extension of the executive anymore. Judges no longer fear falling out of favour with the ruler of the day and we are better off for it. CJ Chaudhry might be retiring and the pendulum of activism might swing back a notch causing desirable correction, but the Supreme Court will remain a powerhouse.

It doesn’t matter that in fighting for the constitutional protection afforded to judges, CJ Chaudhry was in fact fighting for his job. Had he chosen to resign and go home on March 9, 2007, judicial independence would still be in its infancy. His decision to defy the might of the state wasn’t without grave personal consequences. His standing up created a window of opportunity that has strengthened the rule of law.

There is no question that we need a strong independent judiciary to act as an effective check on abuse of power by the executive. What CJ Chaudhry’s reign has done is highlight that we also need an effective system of internal checks to ensure that the judiciary doesn’t commit the misdeeds it is meant to check in others.

Freedom from external interference is one dimension of judicial independence. The other is the ability of the individual judge to discharge functions without interference from peers. This dimension, fettered historically by the misconceived notion of pater familias, has suffered further under CJ Chaudhry who ran a tight ship.

In the initial run after restoration, judges shared the desire to stick together and fight their fights against the PCO judges and the NRO government. This is when we saw large benches and unanimous decisions. The bonhomie probably ended with the challenge to the 18th Amendment.

As some judges refused to become party to striking down a constitutional amendment, the matter had to be sent back to parliament for reconsideration as a compromise.

Internal differences grew with the Memogate controversy and the trigger-happy use of suo motu by CJ Chaudhry. And we saw in the last year excessive reliance on administrative powers to constitute smaller benches, pack off independent-minded judges to other cities along with inconsequential cases and reserve suo motus and other populist matters for Court One.

Both in relation to suo motu and the authority to nominate judges we saw CJ Chaudhry monopolise the collective power of the Supreme Court and the Judicial Commission, respectively. First he managed to get himself granted the exclusive right to make judicial nominations. And then in exercise of judicial power the court diminished the parliamentary committee’s role in scrutinising judicial appointments. While the Supreme Court has rightly ruled that appointments and promotions within the executive must be the product of an open, rigorous and purposive process, if the same principles are applied to judicial appointments under CJ Chaudhry, many of the appointments might not pass muster.

The use of suo motu by CJ Chaudhry has been problematic. Since its use rests on the will of one man, its exercise is random and inconsistent by design. For example, there is no way to understand suo motus over increase in sugar prices or recovery of two bottles of wine from someone’s luggage except as whimsical populism. Media’s role in suo motu incitement also cultivated its raunchy relationship with the CJ office that is not in accord with the judicial code of conduct.

By laying down no clear judicial tests for ‘public importance’ and ‘fundamental rights’ for Article 184(3) purposes or clarifying the nature of relief the court ought to grant, suo motu has become a source of legal uncertainty. The manner of its use denied the accused the presumption of innocence, curbed the right to appeal, and raised doubts about the court ability to act as an impartial arbiter of the law.

The use of suo motu might have cultivated in public mind the image of the chief justice as a saviour. But it has done so at the expense of our ordinary judicial system as everyone now wishes to be heard directly by our highest court. It is true that the need for suo motu arises due to a malfunctioning governance system. But it is equally the failure of ordinary judicial processes that create a need for fire brigade operations fulfilled in turn by Supreme Court’s suo motus.

CJ Chaudhry’s biggest failing is that since June 2005 he has allowed a moribund court system to limp along under his watch. Instead of throwing his weight behind rebuilding and strengthening sustainable judicial processes, he relied on suo motus to create the perception of a functional judicial system, which is nothing more than a top heavy structure suspended in mid air with moth-eaten foundations incapable of serving the judicial needs of all aggrieved Pakistanis.

The moment of utter shame for CJ Chaudhry (and the entire court) was the Arsalan Iftikhar saga. Loud protestations of innocence did not prevent the court’s fall from grace in the pubic eye. If there was a time when CJ Chaudhry ought to have resigned to save his honour instead of showing up for work with the Holy Book in hand, it was when his son was caught red-handed.

CJ Chaudhry’s legacy is a mixed bag. He will be remembered as the politician judge who resurrected an independent judiciary, but driven by power, went too far pushing personal agendas and wading into the business of other vital state institutions. None of this should however prevent the Supreme Court Bar Association from preserving tradition and hosting a farewell dinner for the outgoing chief justice even if not for the man.

The writer is a lawyer.

Twitter: @babar_sattar

SOURCE: www.dawn.com

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The Institute of Optics

News & Events

 May 20, 2013

Institute of Optics shines at commencement festivities

2013 Class with Director Xi-Cheng Zhang
Commencement speaker Steven Chu with Director Zhang
Honorary guest Jie Zhang with Director Xi-Cheng Zhang
Seniors with the Hajim Sign

The Institute of Optics proudly presented its class of seniors who graduated this month. The picture on the right shows Director Xi-Cheng Zhang with (top row from left to right) Anthony Yee, Greg McKay and Hae Won Jung.  Standing on middle step are Sam Steven and James Talerico. Front row is Sarah Walters.

Following is an excerpt from Dr. Zhang's recap about Commencement weekend:
"This past weekend was filled with sunshine and a focus on Optics. It was a privilege to participate in the 163rd College commencement as our undergraduates, graduates, and faculty were honored. Our colleagues, Govind and Miguel, earned the UR distinguished awards for their excellence in graduate and undergraduate teaching, respectively. Two optical physicists highlighted the ceremony: Steven Chu ’70, Nobel Laureate and former US Secretary of Energy, delivered the 163rd College commencement address; and Jie Zhang , President of Shanghai Jiaotong University, received an honorary doctor of science degree. Jie Zhang is the first Chinese to receive such honor in Rochester’s history, and it was rewarding to be present for this historical event. Of the more than 2,300 students who received degrees, eight Optics students earned their BS. Six of these graduated with honors; three with Highest Distinction, one with High Distinction and two with Distinction.  Next year we look forward to doubling this number! Twelve of our students earned an MS, and thirteen achieved their PhD.  Of these new alumni, Greg McKay (BS) and Dennis Fantone (MS) represent second-generation Optics degree recipients. Watching their families celebrate continuing excellence in Optics was heart-warming.  Anticipating the future for each of our newest alumni is awe-inspiring. ...Overall, the weekend was bright with promise as we celebrated achievements, received recognition and launched the class of 2013. Meliora!"


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Old 08-26-2009, 11:23 PM   #3739
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Let's take these SOB's out!!!!!!

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Floyd Little

Originally Posted by bombay View Post
You're spot on about the deterioration of Marriot's voice after about 1972. I saw Humble Pie on every American tour in the '70's, and whether it was the screaming or whatever, he just wasn't as good by Smokin'. He and Frampton were an interesting balance. Kind of lennon/mccartneyish.
Thanks Bombay, I appreciate that.
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Old 12-01-2012, 07:39 PM   #7
Mat'hir Uth Gan
Is It Draft Day Yet?

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I dunno. I do think he is a 1st Round back and would compete with Giovani Bernard to be the first back off the board. Right now, I'd think Green Bay would be very interested around Pick #25.

The guy is 6'0, 220. Runs in the low-mid 4.5s. Is going to have over 1200yds averaging more than 6 yards per carry in the SEC. He also has decent hands. And he's only a Junior in a timeshare with Yeldon whom is a future Top 10 back, or he'd have massive numbers. He was really good spelling Richardson last year as well and has substantial similarities to Trent.

If he's not a 1st Round back, I'm not sure what is.
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Old 02-28-2013, 10:40 AM   #19

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With the 18th pick in the 2013 OM draft the Dallas Cowboys select....

DT Johnathan Hankins from THE Ohio State University - 6'3" 322

Ohio State's great 2012 season could be attributed to many factors; one big reason for their success is DT Johnathan Hankins. The 6'3", 332 lb anchor of their defensive line is a true difference maker.
Hankins is best-suited as a RDT or NT that can occupy double teams and control the middle of the trenches. Although he plays with inconsistent technique, he is powerful enough to make up for it. He holds his ground well and does not get pushed around by blockers. One adjustment he will need to make is keeping his pad level low and driving with his legs rather than attempting to just manhandle players with his upper body strength.

With the recent drunk driving DT's that the Dallas Cowboys currently employ it's time to start thinking about building their future at that position. Johnathan Hankins is the perfect fit to do that. Monte Kiffin is going to build this defense from the inside out, and front to back. Johnathan Hankins has both the size and strength to be able to do that. Let's send it over to Jon Gruden to get his take on this pick... Jon?

Gruden: "This guy Johnathan Hankins... let me tell you something man, he is a true FORCE to be reckoned with at 6 foot 3 inches 322 pounds he is going to be knocking the grins right off of opposing linemen and running backs... I LOVE this pick for the Dallas Cowboys."

Pick was made at 10:39
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1. Q: What are qSTAR qPCR Primer Pairs and panels?
A: qSTAR qPCR Primer Pairs are pre-designed, qPCR tested and ready-to-use primer sets for SYBR Green based qPCR experiments. The primer pairs are also available in a 96-well panel format covering broad range of pathways for cancer biomarker profiling.
2. Q: Can qSTAR qPCR Primer Pairs be used in probe-based qPCR?
A: No. This kit has not been designed to accommodate any commercial probe based qPCR.
3. Q: What is the Tm of the primer and what is a typical size of the amplicon?
A: The Tm of a qSTAR qPCR Primer is around 60, and the amplicon is around 95 to 140 bp.
4. Q: Does a qSTAR qPCR Primer Pair amplify an exon junction in a cDNA target?
A: Whenever possible, an amplicon by a qSTAR qPCR Primer Pair covers exon junction or junctions.
5. Q: Primer Pairs and Standards
A: My qPCR machine is not on the list of compatible machines on the website. Do you provide qPCR primer panels in plates compatible with other machines?;Yes, we can provide our primer panels in other plates within approximately one week of receiving the custom order. Please email the catalog numbers of the qPCR primer panels needed plus the catalog number and manufacturer of the plates to techsupport@origene.com. Our technical support scientists will prepare a custom quote that can be used to order custom panels.
6. Q: If I provide a list of genes I want to assay, can OriGene prepare a custom qPCR primer panel?
A: Yes, OriGene can design a custom qPCR primer panel. Custom panels are offered for human and mouse genes in 200 reactions format for purchase, delivered in 2-D bar code matrix tubes.
7. Q: Where can I find the sequence of the primers in each panel?
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8. Q: How much primer pair is in each well?
A: Each well in 200 reactions format contains 2 nmol of lyophilized primer pair. Please re-dissolve in 200ul of water so the final concentration is 10uM. Each well in the PCR plate contains 10pmol of primer pairs. The primers will be re-suspended once the PCR reagents are added to the plate.
9. Q: What is a qPCR template standard?
A: A gene specific qPCR template standard is a tube of linear DNA made from a full-length cDNA plasmid. The copy number of a template standard solution is determined by the PicoGreen method and calculation based on MW.
10. Q: Can a template standard be used in a probe-based qPCR?
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11. Q: In your protocol, it is recommended to make six serial dilutions with one log of difference, can I make more dilutions and with different dilution scheme?
A: Yes, as long as it is diluted in the qPCR detection range and each dilution is mixed thoroughly.
12. Q: Can I use my own buffer to dilute the template standard instead of the buffer in the kit?
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13. Q: Does OriGene offer custom-made template standards?
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14. Q: What is the OriGene guarantee on qPCR primers and template standards?
A: OriGene qPCR primers and templates are warranted for SYBR Green qPCR experiments. If your experimental results are not satisfactory, our scientists will work with you to pinpoint the problem and, if it is determined that our products are at fault, OriGene will refund your money in the form of a credit.
15. Q: How should I calculate the copy number of my gene of interest?
  1. If the cDNA templates in your samples are single-stranded such as cDNA from RT reactions, theactual copy number of your gene of interest is two times the number you got by comparing to theqPCR standards. For example, if the copy number of your gene is 5 copy/-ª+îl from the standardcurve of a qPCR program, the actual number is 10 copy/-ª+îl. The reason is that the first cycle for asingle-stranded sample is to make the complementary strand; therefore, there is one cycle delayin the PCR reaction compared to the double-stranded cDNA template standard's reaction.
  2. If the templates in your samples are double-stranded, the copy number of your gene of interest isthe same as that calculated according to the qPCR standards.
16. Q: I am writing a paper for publication and need to describe this product. How should I cite?
A: We recommend that you refer to the product by its specific catalog number and refer to us as OriGene Technologies (Rockville, MD). Furthermore, we'd love to hear from you when your paper is published. Inform us and we will send a gift.

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'The Voice' recap: Episode 4

Man, those judges on “The Voice” did some major work tonight, scooping up more talented hopefuls as we endured (enjoyed?) another week of blind auditions. I seem to have forgotten that this show is a marathon, not a sprint – but we’re so close to finishing the first stretch of the race.

Next week’s the last week of the blind auditions. Then we get to the battle rounds. And just in case you’re thinking this show takes itself too seriously (which it does), I offer the continued appearance of Cee Lo Green’s cat, Purrfect, as whimsical balance.

Team Christina

Jonathas, a Brazilian-born crooner who taught himself English by listening to music. His take on Usher’s “U Got It Bad” had Blake whispering “He’s gotta be good lookin’” and Cee Lo proclaiming that he really did think it was Usher singing -- that’s how good Jonathas was. Agree to disagree, Cee Lo.

Monique Benabou, 23, who sang “Mr. Know It All” and is in general, a big bundle of curls and curves and adorableness.

Anthony Evans, 33, from Dallas. He’s the son of a preacher-man (I just wanted to type that, I’m sorry) and had struggled in the past between defining himself as a gospel singer or as a contemporary Christian musician. His take on Marvin Gaye’s “What’s Going On” makes it seem that he could easily do both.

Ashley De La Rosa. Aside from her awesome name and equally awesome take on “Shark in the Water,” we don’t know too much about this one yet. We’re not free of the clipped montages of winners and losers just yet.

Ten down on Team Christina, two to go.

Team Blake

Street performer Naia Kete, 21 and dreadlocked (the entire family’s dreadlocked – mom, younger brother, everyone, and it’s just lovely). Her cover of Bruno Mars’ “The Lazy Song” had Blake and Cee Lo turn, with Christina noting that the texture of Naia’s voice is remarkably similar to last year’s runner-up (and Team Blake contestant) Dia Frampton.

Charlotte Sometimes, 23, from New Jersey. Her sob story was something along the lines of “I had a jaw disease and didn’t think I would sing again,” which is just bizarre. And Charlotte “Sometimes”? Who are you the rest of the time? But goodness, is her voice odd and lovely.

Jordan Rager, again a victim of clipped editing and brief montages. I can’t remember a darn thing about his performance, other than it happened, but Blake called him “real country.”

And another edited-down performance of Pink’s “Just Like a Pill” by someone named ALyX who I’m sure is all kinds of cool and edgy.

Only two more spots to fill on Team Blake.

Team Cee Lo

Tony Vincent, 38, and a Broadway veteran whose credits include stagings of “Rent,” “Jesus Christ Superstar” and “American Idiot.” He’s worked extensively with Queen, so the cover of “We are the Champions” wasn’t too much of a surprise.

Jamie Lono, who makes sandwiches at Potbellys and who just looks absolutely flabbergasted he’s even on the show. His take on “Folsom Prison Blues” had Adam’s chair spinning before Jamie could even really get into it. Elbows tucked into his sides, both hands gripping the mic, Jamie just says “I make sandwiches for a living.” While Adam promised him “I’ll make sure you never have to make a sandwich again,” he went with Cee Lo. Who clearly eats a lot of sandwiches, so I’m sure that’ll be a good match.

Justin Hopkins, 30, who used to be in the Carson Daly House Band. A) Well, we all know how he got onto this show. B) In what lifetime did Carson Daly have a house band? I forgave the Carson Daly connection, however, the minute Justin started singing David Gray’s “Babylon.”

Three slots still open on Team Cee Lo.

Team Adam

Nicolle Galyon, from Nashville, wants to be the first “piano girl” in country music. She got a little … hmm, breathless? when Adam turned his chair, prompting Blake to tease him later: “No, man, I understand it. You make me giddy. You make me fall apart.” The bromance these two have makes the world a better place.

Karla Davis. Another country girl from Nashville, and another victim of the too-short performance. Her cover of “If I Die Young” sounded pretty charming, though.

Mathai, 18, from Dallas. This is why this show is awesome. Tiny little unassuming people with awesome voices. Mathai’s entire family consists of either doctors or nurses, and she laments that her mother has a tough time explaining that her daughter isn’t in school, doesn’t have a career, etc.  I wouldn’t worry too much, Mom. Your kid’s got a set of pipes. I mean, she covered Adele. That’s tough.

Two more spots on Team Adam.

Leftover Thoughts

Dan Auerbach doesn’t dance when he sings “Tighten Up.” Which means if you plan on auditioning with a Black Keys song, you shouldn’t either.

At no point, if auditioning, should you say, “I need this to happen.” You won’t make the cut, and all that footage will do is give the editors the power of foreshadowing. Don’t give them that power, blind auditioners. You know better.

Copyright © 2017, Orlando Sentinel


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Linux We all know Synaptics, the company that seems to produce just about every touchpad you can get your hands fingers on. Their touchpads also do a lot of multitouch and gesture stuff, but up until now, their set of gestures, the Synaptics Gesture Suite, was only available on Windows. Luckily, they've ported it over to Linux, and made it available for OEMs building Linux laptops.
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Gone fishing
Member since:

.... Or worse, they have some Trojan or malfeatures in their code they don't want you to know about.

In any case, it's probably wise to avoid putting yourself at risk by running such closed software on your computer.

I can see the argument against binary blobs is a good one, but this is over stating the case, hardware vendors supporting Linux is a good thing, OK it would be better if this was with opensource drivers, but it is still a step in the right direction.

I wouldn't suggest installing this on your server, but on a less than mission critical netbook, smartbook etc it will be a good thing.

So - Year of the Linux netbook, smartbook, laptop! I think year of the Linux Desktop has already happened just many people haven't noticed.

Reply Parent Score: 5


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Windows As of today, Microsoft won't allow manufacturers to install XP on new netbooks," says blogger Kevin Fogarty. "That doesn't mean corporate customers who special-order hardware with XP won't be able to get it, or even that its market share ( 60 percent!) will drop any time soon.... It just means XP has taken the first babystep toward obsolescence and the long (really long, considering its market share) slide down toward the pit of minor operating systems like the MacOS X (4.39 percent) , Java ME (.95 percent) and "Other" (which I think is an alternative spelling for "Linux" (.85 percent).
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Aiming at the total user experience with quasi-dictatorial control of the final hardware and software appear to be what makes Apple successful in introducing its new devices. The X86 based OEMs have been forced to focus on only the hardware aspect of their devices as the software was handled by Microsoft. Applying the same development model with a minor adaption of a Linux distribution to the device ended-up with the bad results you mentioned.

A few comments about ARM based netbooks (and netpads) have been made.

What I am hoping, as an user, is for a major manufacturer to develop a device based on this platform plus a cleaned-up and modernized Risc OS (please keep the OS in ROMs for warp-speed boot!) and web-related apps along with some sound and video manipulation capabilities.

Reply Parent Score: 1


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IconPeople shouldn't read anything into the fact that of the three laptops set up for last week's 'PWN to OWN' hack challenge, the only one left standing was running Linux, said the security expert who oversaw the contest. "There was just no interest in Ubuntu," said Terri Forslof, manager of security response at 3Com Corp.'s TippingPoint subsidiary, which put up the cash prizes awarded at the contest last week at CanSecWest. "A contest such as this is not a measure of relative security between operating systems. It's not an accurate barometer."
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Technology White Papers

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Opera Software Opera have announced the general availability of Opera 10 beta. Opera 10 includes an improved rendering engine Presto v2.2. The beta adds a new default skin and a couple of new features, notably "Turbo", a proxy compressor for dial-up users, and tab previews. The result? Complete fail. Read More for why and a quick screenshot tour. addendum: As an apology to the community for the reckless and inadequate review I will be doing it again, properly, taking into consideration your fine comments.
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RE[2]: I like it
by pandronic on Thu 4th Jun 2009 05:27 UTC in reply to "RE: I like it"
Member since:

I think it looks great. It seems to integrate quite well in OS X. So what if it's a little purple and black? Will the mighty Apple gods be pissed because Opera didn't follow their precious HIG by the letter?

Reply Parent Score: 4


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Multimedia, AV Yes, I broke my own rules and used a "breaking" modifier for this story (let me have my fun for once). Here we have it, as the rumour mill suggested, Google has released the On2 VP8 video codec as open source (royalty free, BSD-style), while also launching the WebM container format which combines a VP8 video stream with Vorbis audio. Support for WebM has been enabled on YouTube's HTML5 beta, and you can download patches against ffmpeg as well as DirectShow filters for Windows (Gstreamer plugins are labelled as "coming soon"). Mac users are out of luck for now; no QuickTime plugins have been announced yet. Update: The WebM blog is now open - and the list of partners is pretty decent already. It includes ARM, NVIDIA, AMD, Qualcomm, and many others. Update II: VP8 will be baked into Flash. Update III: The Opera labs version with WebM support has been released too, for Linux, Mac, and Windows.
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RE[2]: Awesome news.
by DigitalAxis on Wed 19th May 2010 20:57 UTC in reply to "RE: Awesome news."
Member since:

Hopefully, if Froyo will be able to support Flash/h264 video on current devices, Gingerbread will be able to support WebM on those same current devices.

I'd hate to spring for an Android handset now and find out I'm crippled due to lack of hardware acceleration.

Reply Parent Score: 2


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Hair Color Trends and Tips for 2011

2011 is coming up, so you need to get everything down pat in order to stay in vogue! That’s why we decided to focus your attention on hair color trends for 2011 as you know hair color can play a very significant role when it comes to changing your look. For perfect hair color one should match it to the skin tone and eyes perfectly so our hair color trends and tips for 2011 will definitely help you to choose a hair color for your new look to enhance your natural beauty.

Share |

Who doesn’t dream of a luxury hair color that accentuates beauty of the hair enhancing the complexion and facial features? And hallelujah, nowadays that’s possible to get as the hair dye makers have developed a vast palette of hair dye shades just so they can suit every complexion and all the latest fashion tendencies.

Hair Color Trends and Tips for 2011

Choosing right hair color is not an easy task, but with our hair color tips you’ll definitely find the perfect hair color for you. Follow hair color trends 2011 and turn to a professional stylist for best results.

One thing you should learn is naturalness as it is an absolute key word in fashion for 2011 and when it comes to choosing a hair color you are to be guided by naturalness as well in order to be trendy in the forthcoming 2011 year.

Red hair color is an absolute trend for 2011! This hair color is a perfect choice for freckled women to get an adorable look. You can opt for a more natural look or go dramatic with flashy hair coloring. But be very careful as uber bright red hair color will highlight all the possible flaws on your face.

Hair Color Trends and Tips for 2011

Blonde hair color is another one that is uber- popular alternate for 2011 as we know that blonde hair has a certain zest which enhances the sex appeal. There is such a great variety of blonde shades to choose from and I must admit 2011 is that year when you can get any shade. It means you will be in vogue irrespective of the blonde hair color selected.

Hair Color Trends and Tips for 2011

As for brown hair color, it suits most skin tones so they are always utterly popular and the 2011 is no exception. There are a great variety of brown hair color shades to choose from depending on what suits your complexion, from dark chocolate brown to light caramel. If you have swarthy or a little bit tanned skin complexion go for a wonderful coffee or taffy brown hair color.

Hair Color Trends and Tips for 2011

And finally black that is a permanent trend when it comes to hair color. In 2011 black is a great alternate for women whose complexion is pale as well as a bit tanned. But be very carefully as this hair color doesn’t suit everybody. But if you have fair skin tone and blue eye, don’t hesitate, black hair color would perfectly suit you!

Hair Color Trends and Tips for 2011

And the only thing you should adopt that regardless to hair color you’ve chosen (even if it perfectly suits you) all the efforts would be vain without a proper hair care!

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4 Responses to “Hair Color Trends and Tips for 2011”
  1. Claire Says:

    Great article!

  2. health Says:

    thanks you nice very good

  3. Anonymous Says:

    great hairstyles

  4. Lekha Says:

    Wow…….its cool

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Overclock.net › Forums › Components › Sound Cards and Computer Audio › Fidelio X2 after burn in give rolling and crackly
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Fidelio X2 after burn in give rolling and crackly

post #1 of 4
Thread Starter
After a bit of burn in my X2 being to have an highter volume in the bass compared than treble and bass is rolling
im using the integrated 1150 now, but also with notebook and smartphone its the same.

the general volume is high.
I test it with game and MP3 320
post #2 of 4
Sorry to hear you're not enjoying them right now. There is no question about it, the X2's have a punchy bass. I suppose this is what reviewers are referring to when they describe it as a fun headphone. It's very dynamic and plays well with music that is similar. There are a couple of things to check first with your computers. Make sure all the Realtek enhancements are turned off. There are a lot of them and their descriptions are vague or non-existent. The X2 doesn't need any boosting in those areas and doing so could contribute to the sound you're describing. The X2's are 30 Ohm, so they are going to come off loud when paired with a high gain source, like a Sound Blaster Z, etc. Even the newer MB's have 600 ohm headphone amplifiers (20% volume for me), so your just need to use the low end of the volume scale. The smartphone will not have that issue.

While typing this, I have been listening to some Deep House tracks to try and create the situation you're describing. With my external DAC/Amp combo, I am not really hearing it. The bass is quite evident as you would expect, but it's not muddy or drowning out the other end of the spectrum. Moving the plug over to my Asus X99 on board (Realtek 1150), a lot of grain becomes quite evident. The bass does not sound as sharp and there are all kinds of things in the midrange that are making me unhappy. It's not a fair comparison between my external set up and a motherboard that in it's entirety, costs less, but that is the reality of it. THe X2's are sensitive enough to reveal other weak links. My X2's have about 70 hours of use so far.

You can try playing around a bit with the Realtek codecs if they were already off. In the DTS Ultra PC II section, try spectral enhancement with the bass off and the treble a few notches up. Normally, I don't like this kind of thing, but it's really user preference. Also, read through the review below. I am sure you've already read it, but take note of the changes he highlights after a 100 hour break-in. It may ease your concerns somewhat, or help you make a decision in a different direction.

post #3 of 4
Thread Starter
im using realtek ALC1150 supremeFX III.

at first the bass was not so loud, now the volume during bass is high compared to treble, for example during song if you set the volume around the bass you will hear the voice too weak .............. instead if you set the volume around the voice, when bass r there your ear die ( and the end of bass effect was bad )

What amp/dac combo are u using ?
post #4 of 4
I used it with my Woo WA7 for a week or so before finding a permanent solution for the PC in A Creative X7. But I also used with my trusty old Soundblaster Z on the first day right out of the box. I thought it sounded fantastic on the Z on day one, so I don't think you need to be too particular with your AMP/DAC selection.

There is another thing to check. Make sure the X2 cable isn't twisted, bent, or running around something carrying a current. I haven't had any real issues, but did notice a little crackle when moving the cable around a lot in the beginning.

**In fact, I don't like the cable much at all. I switched it out for a cheap $8 one I had from when I had the X1. I didn't notice any difference on an iPhone, but through the Creative X7 it now sounds far more vivid and clear in one game where I didn't like using the X2. Limited testing. I'm not offering this as evidence toward the benefits of cable swapping, but simply as trouble shooting. If you have a cheap 3.5 to 3.5mm cable around, or if it's worth $8 to you, pick one up and see if some of the problems don't go away.
Edited by c-attack - 1/14/15 at 2:14pm
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Spike TV Video Game Awards Wrap Up

Category: Gaming
Posted: 09:45AM

Spike TV held its annual awards show last night, giving out awards for the best games. Among the big winners were Red Dead Redemption for Game of the Year and BioWare for Studio of the Year. A full list of winners and awards can be found here. However, the bigger news of the night came in the form of game reveals and trailers. A number of games were confirmed, while many others were given an official release date. The Elder Scrolls V: Skyrim, Prototype 2, and SSX: Deadly Descents were confirmed by their respective publishers. Uncharted 3: Drake's Deception and Mass Effect 3 were given release dates taking place during the holiday season next year. Forza 4 and Resistance 3 will see the light in the Fall of next year. Additionally, Professor Hugo Strange was revealed as a villain in Batman: Arkham City.

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Andrewr05 on December 12, 2010 12:30PM
I would rather have watched the Oscars than this drivel...
CheeseMan42 on December 12, 2010 02:15PM
That's why you just wait to see the results and trailers. :P

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Definition of misuse noun from the Oxford Advanced Learner's Dictionary

BrE BrE//ˌmɪsˈjuːs//
; NAmE NAmE//ˌmɪsˈjuːs//
[uncountable, countable, usually singular] (formal) the act of using something in a dishonest way or for the wrong purpose synonym abuse alcohol/drug misuse the misuse of power/authority a misuse of public funds
Extra examples Can manufacturers be held liable for the criminal misuse of their products? He was accused of the misuse of public funds. He was fired for computer misuse. alcohol misuse among teenagers causing pollution through misuse of pesticides the misuse of power/authority
See the Oxford Advanced American Dictionary entry: misuse

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stereo noun definition in Radio technology topic from the Oxford Advanced Learner's Dictionary

 noun: Radio technology topic
(also stereo system) [countable] a machine that plays CDs, etc., sometimes with a radio, that has two separate speakers so that you hear different sounds from each a car/personal stereo Let's put some music on the stereo.

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The media


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Quote Originally Posted by naptownmenace View Post
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Anyone who compares Reggie's regular season performance to any other SG is totally missing what made him a HOFer. The thing that set Reggie apart was his ability to come up big in the playoffs. Reggie averaged 21.1 ppg in the playoffs. Before his role deminished during his last 3 seasons, his average was a gaudy 24 ppg - nearly 5 points more than what he averaged during the regular season!

Mitch Richmond never made any noise in the playoffs or in the clutch. A lot of that is due to his poor level of talent around him but his play during the playoffs never matched his during the season production.

Tracy McGrady was one of the best regular season SGs of all-time. He couldn't get his teams, even with a 3-1 lead with Orlando or an in his prime Yao Ming and Steve Francis, to the 2nd round.

Glenn Rice was a great scorer for about 2-3 years. Ultimately he fizzled out due to injuries even though he was a very good player that torched the Pacers a few times during the 2000 Finals.

Reggie was clutch. All of the NBA greats agree. That's what made him famous, hence he is in the Hall of Fame.

Right. Reggie won playoff games and series with more regularity than players that were better than him. Those playoff wins were team accomplishments, not individual accomplishments. Just as Russell's championships don't make him a better player than Wilt just because Russell had a better supporting cast of HoFers. Reggie had longevity and playoff wins and thus he's remembered more favorably than players that were better than him. That's why I think top-ten is appopriate but not top-three or top-five.


text:
 Household Dust

Household Dust

Declare War on Dust

Dust is a real problem for every household. Worse still, dust seems to be a growing problem. If that’s how you feel, it’s not your mind playing tricks on you. Weather patterns are more extreme with winds bringing more and more dust. Houses are increasingly more energy efficient, making them dust traps that collect and hold the dust. And, in this electronic age, we are putting more and more electrical elements into our homes, and all that heat and electricity pulls and draws dust like nothing else can. So, if you thought your house is getting dustier, you are not imagining it. It is getting worse.

So, what can you do to stop the deluge of dust? Fortunately, there are some steps you can take to help keep dust down and lessen your housework – and maybe even help yourself breathe a little easier. Because all that dust takes a toll on our health and it's just plain dirty.

To curtail dust, pay special attention to these areas:

Doormats. Invest in good quality doormats inside and outside the doors. This can stop a lot of the dust from ever entering your home.

Remove shoes. Just by removing shoes at the door before entering the house, you will cut down the amount of dirt, germs, and pollutants from entering the house and carpets will last longer.

A/C filters. Use good quality air conditioning and heating filters and change them regularly.

Check seals. Doors and windows should be checked to make sure seals are working well and have not deteriorated. You should also check seals to the basement and attic to make sure the access to these areas seals tightly and that you aren’t pulling in squirrel droppings from the attic every time you open the front door.

Check the dryer. Sometimes dryer vents are filled with lint causing them to pour lint into the air instead of venting out the house. Check the hose to make sure it’s clear and blocked. Make sure you pull the lint trap out of the dryer and clean it with each and every load.

Replace carpet in traffic areas. If you have carpeting at any door openings, you have a dust trap. Consider replacing entry and traffic walkway areas of your home with hard surface flooring that won’t attract and trap dust.

Have A/C ducts cleaned. Every other year or so it's a good idea to have A/C and furnace ducting cleaned to remove any molds, insulation, or dust.

Wash plants. Make sure plants are kept clean by washing them in the shower or yard. Both real and silk plants should be washed at least once a year to remove dust and keep them clean.

Pets. If you have indoor pets, you have animals losing hair and skin cells in your house all day long – something dust mites love to eat. This makes vacuuming especially important. People with indoor pets should vacuum three times a week to keep dust down and pay special attention to vacuuming upholstered furniture.

Dust walls. Yes, dust walls – especially those surrounding electronics or TVs. With media rooms becoming more and more popular, the heat and electricity pull dust in around the appliances. Use a Swiffer Max (extra large Swiffer; see below) to “mop” the walls of dust.

Wipe as you go. Keep a damp towel with you to wipe down baseboards, handrails, sills, and door tops as often as possible as you are cleaning your house.

Use the whole vacuum. While this may seem silly, most vacuums are like icebergs: only 1/10 of them is appreciated. Most of the time, the vacuum is run down the middle of the floor and that’s it. However, the best part of the vacuum is usually with all the attachments. Use the edge and crevice tool to get in between and under and to run down the edge where the wall meets the floor – a serious dust collection area. You should also use the brush to vacuum electronics, blinds, and lampshades, the upholstery tool to go over mattresses, drapes, and upholstered furniture.

Vacuum regularly. Once a week is a minimum and twice a week is excellent.

Change the vacuum bag/empty the canister. A full vacuum bag has no suction power and won’t pick up as much dirt and dust. Change your bag when it is more than half full. If you have a canister on your vacuum, empty every time you vacuum. Why leave that dirt in there if you don’t have to?

Add landscaping. If you have any dirt or dry patches in the yard, cover these over with grass, plantings, or mulch. Blowing dirt is a big source of dust. Your vacuum is one of the best defenses against dust Nothing can replace the value of a good quality vacuum in the war on dust.

A good quality vacuum will provide you with the suction and onboard tools you need to get the dusting and cleaning job done. An old, poorly maintained vacuum, with an over-filled bag is no help to anyone and can actually put dust into the air, making the house dirtier, not cleaner. Make sure you have extra bags or that you empty the dust collection cup each and every use.

My personal favorite are Dyson vacuums. These vacuums don’t lose suction and have tools that are actually easy to use, extra long hoses and cords, and no bags required. The ball vacuum allows you to turn off the beater brush with the touch of a button. They have heavy HEPA filters and don’t require you to buy bags (which are big suction reducers). In short, if you need a new vacuum, this should be the one you buy. I personally recommend them.

Household Dust -- What It is, Where It Comes From, and What to Do About It