Posts Tagged ‘Protocols’

The bacterial industrial revolution

 :: Posted by American Biotechnologist on 08-31-2011

George Church, the talented genetic professor, has made headlines once again. We are very fond of George Church and have written about him and his work several times in the past (see: George Church: The Father of Personalized Genomics, New tools for rewriting the code of life and A Scientific Legend’s Approach to Solving Problems and Developing Technologies).

The latest article, appearing in The Boston Globe, talks about Dr. Church’s approach to synthetic biology and his “broad brush” approach of editing bacterial genomes to devise powerful new technologies.

To read more click here. H/T to Genomeweb for the find.

Still stuck on tank or semi-dry protein blotting?

 :: Posted by American Biotechnologist on 08-17-2011

Learn why the Trans Blot Turbo is faster than tank and semi-dry blotting techniques.

Bio-Rad Video 5/11/11 from Lab Manager Magazine on Vimeo.

How many reference genes do you use in qPCR?

 :: Posted by American Biotechnologist on 08-10-2011

In the past, we’ve discussed the importance of selecting appropriate reference genes for your qPCR experiment (also see point 7 of the MIQE guideline checklist). This means that it is important to select genes that do NOT exhibit any changes in expression under the treatment conditions you are studying. This is easier said than done!

“Once upon a time” everyone used either beta actin, 18s, or gapdh as reference genes. Their expression never changes, right? Wrong! So which genes should you choose? If you try to figure it out using previous papers, how do you know that they’ve chosen the correct genes? If you run a few genes side-by-side and try to compare their expression both under treatment and control, which one should you set as the baseline and which one can you say is for sure moving (it’s all relative isn’t it)?

One of my twitter friends told me that she uses six reference genes in her qPCR experiments. I used to use two. That got me thinking…how many reference genes does the “average” lab use? Please help satisfy my curiosity by participating in the poll below!

How many reference genes are you using in your qPCR experiment?

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7 Great Resources for HRM Analysis

 :: Posted by American Biotechnologist on 08-04-2011

Bio-Rad Laboratories recently launched the Precision Melt Supermix, which is a high-perfomance supermix for both genotyping and epigenetic analyses.

In honor of this launch, we invite you to review some of the resources (including technical notes, review articles and video tutorials) that we have posted on high resolution melt analysis. Feel free to to click on any of the links below for further details:

Brilliant yet simple method for labeling polyacrylamide gels

 :: Posted by American Biotechnologist on 03-24-2011

Most people who have run polyacrylamide gels have at one point or another gotten confused with the gel’s orientation. This is especially true following protein transfer from the gel to a membrane for the purpose of western blotting or other downstream processing. This can be extremely frustrating and may even jeopardize your entire experiment if you are unable to tell the right side of the gel from the left or your control samples from your treatment group.

Several methods have been created to help bench scientists avoid this problem. These include the use of multicolored protein ladders and marking a predefined corner of your membrane once the protein has been transfered (I cut the bottom left corner of the membrane).

Today, Arefeh Seyedarabiclose from the Department of Structural and Molecular Biology, University College London published a new and very basic method for labeling polyacrylamide gels on the Nature Protocol Exchange website. Essentially, Arefeh suggests labeling the bottom inside corner of the long glass plate (facing the gel) with permanent marker). During electrophoresis the label will transfer from the plate to the gel, thereby permanently labeling your gel. Simple and brilliant!

To see the full method and associated figures click on A method for labeling polyacrylamide gels.

If you would like to share other great molecular biology methods, please post a comment or send us an email using the email button at the top of the page.

Reference:
Arefeh Seyedarabi, A method for labeling polyacrylamide gels, Protocol Exchange (2011) doi:10.1038/protex.2011.222, Published online 24 March 2011