Posts Tagged ‘Real-Time PCR’

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:

The MIQE iPhone App

 :: Posted by American Biotechnologist on 07-25-2011

We have written many posts about the MIQE real time PCR standards that are basic requirements for anyone engaged in real time PCR experimets. Now there is a new tool for all ipod/iphone users to add to their arsenal. A MIQE qPCR app!

The MIQE app helps scientitst review scientific work and check their own project’s MIQE compliance. Plus, the app includes a list of the most current qPCR news and events and “emergency” contact numbers that you can call/email should you have any questions about your qPCR experiments.

The application was developed by Dr. Afif Abdel Nour, Associate Professor in Nutrigenomics at LaSalle Beauvais, in collaboration with Dr. Michael Pfaffl and was sponsored by Bio-Rad Laboratories .

The Real-Time PCR iPhone App

 :: Posted by American Biotechnologist on 07-25-2011

An easy-to-use qPCR resource, Bio-Rad’s Real-Time PCR iPhone application includes the Real-Time PCR Applications Guide for researchers who want to learn more about designing, analyzing, and optimizing real-time PCR experiments. Another feature is the qPCR Doctor, an interactive troubleshooting tool for resolving problems relating to real-time PCR assays. The Real-Time PCR iPhone Application also includes a qPCR Assay Design section which provides guidance for designing a qPCR assay, information on validating and optimizing your qPCR assay, and different methods for analyzing qPCR data. This application puts three of Bio-Rad’s best real-time PCR resources at your fingertips.

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Amplification Central: A Hidden Gem for the Molecular Biologist

 :: Posted by American Biotechnologist on 01-27-2011

One of the internet’s hidden gems for molecular biologists is the amplification central gateway by Bio-Rad Laboratories.
The gateway includes a ton of resources for those involved in designing and executing both real time and conventional PCR experiments. There’s a PCR Doctor section which includes a real time PCR doctor for diagnosing and treating real time PCR trouble, and a conventional PCR Doctor for diagnosing PCR trouble after the amplification product is run on an agarose (or PAGE) gel.

The site also has tutorials on Real-Time PCR Fundamentals and Real-Time PCR Chemistries which include downloadable presentations and an Assay design section that includes Primer and PCR Product Design, Assay Validation and Optimization Overview and Quantitative PCR Analysis.

To top it all off, the organizers have included a tip of the week (this week’s tip is: Using a standard curve on two different plates can serve as an inter-run calibrator) which should encourage you to visit the gateway regularly.

Click here to visit amplification central today!

If you know of other hidden internet gems for molecular biologists, please share!