Posts Tagged ‘qPCR’

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:

Tools for Epigenetic Chromatin Analysis

 :: Posted by American Biotechnologist on 07-27-2011

In this slideshow, you will learn the latest epigenetic techniques including:

  • discriminating epigenetically inactive chromatin from active chromatin
  • discriminating between aberrant and Monoallelic DNA methylation
  • predicting gene expression levels via chromatin structure assay
  • analyzing how DNA methylation affects promoter activity

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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Advancing qPCR Bio-Rad Style

 :: Posted by American Biotechnologist on 07-06-2011

Bio-Rad Laboratories has just launched a brand new qPCR supermix which has significant advantages over other technologies on the market. The SsoAdvanced SYBR Green Supermix is based on Bio-Rad’s patented Sso7d fusion protein technology. The dsDNA binding protein, Sso7d, stabilizes the polymerase-template complex, providing superior inhibitor tolerance, increased processibity and greater speed.

With Sso Advanced SYBR Green Supermx you can expect to:

  • Acheive superior gene expression results under various cycling conditions
  • Increase sensitivity and efficiency of detection from compromised samples
  • Decrease run times and time to results without compromising on qPCR data quality
  • Obtain better results with predeveloped qPCR assays

For more information and experimental data download the SsoAdvanced SYBR Green supermix product sheet.

To learn more about Bio-Rad’s qPCR solutions visit www.bio-rad.com/qpcr