Correction of RT–qPCR Data for Genomic DNA-derived Signals with ValidPrime
News Jan 09, 2012
Genomic DNA (gDNA) contamination is an inherent problem during RNA purification that can lead to non-specific amplification and aberrant results in reverse transcription quantitative PCR (RT—qPCR).
ValidPrime measures the gDNA contribution using an optimized gDNA-specific ValidPrime assay (VPA) and gDNA reference sample(s). The VPA, targeting a nontranscribed locus, is used to measure the gDNA contents in RT(+) samples and the gDNA reference is used to normalize for GOI-specific differences in gDNA sensitivity.
This article demonstrates that the RNA-derived component of the signal can be accurately estimated and deduced from the total signal. ValidPrime corrects with high precision for both exogenous (spiked) and endogenous gDNA, contributing _60% of the total signal, whereas substantially reducing the number of required qPCR control reactions. In conclusion, ValidPrime offers a cost-efficient alternative to RT(_) controls and accurately corrects for signals derived from gDNA in RT–qPCR.
To read the full article, please click on the link provided.
A human pluripotent stem cell line has been engineered which contains two ‘suicide genes’ that induce cell death in all but the desired insulin-producing cells. This double fail-safe approach opens the door to creating safe cell-replacement therapies for people living with type 1 diabetes.READ MORE