:: Posted by American Biotechnologist on 08-24-2011
Some of the technical challenges in qPCR research can be traced back to the reverse transcription step and the synthesis of cDNA for qPCR analysis.
Limitations in the maximum amount of template RNA that can be added to an RT reaction make it tougher to detect low-abundant target genes and limits the amount of data that can be obtained from a typical 20ul RT reaction.
Bio-Rad Laboratories has recently launched the iScript advanced cDNA sythesis kit for RT-qPCR as an answer to some of these challenges.
iScript advanced is a 2-tube format cDNA synthesis kit with a short protocol (35 minutes) and an allowed RNA input amount of up to 7.5ug.
The kit will:
- increase the amount of data that can be obtained from an RT reaction (great for high-throughput labs)
- enable you to analyze a larger number of target genes from a single sample
- increase the sensitivity for certain target genes due to higher RNA input capacity
- cover a broad dyamic range and sensitivity
For more information on iScript Advanced including experimental data download the iScript Advanced product bulletin.
To read more about Bio-Rad’s qPCR solutions visit www.bio-rad.com/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!
:: 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:
:: 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
:: 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.