Correction of RT-qPCR data for genomic DNA-derived signals with ValidPrime

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F12%3A00379016" target="_blank" >RIV/86652036:_____/12:00379016 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.1093/nar/gkr1259" target="_blank" >http://dx.doi.org/10.1093/nar/gkr1259</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1093/nar/gkr1259" target="_blank" >10.1093/nar/gkr1259</a>

Result language
angličtina
Original language name
Correction of RT-qPCR data for genomic DNA-derived signals with ValidPrime
Original language description
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). Currently, there is no alternative to RT(-) controls toevaluate the impact of the gDNA background on RT-PCR data. We propose a novel method (ValidPrime) that is more accurate than traditional RT(-) controls to test qPCR assays with respect to their sensitivity toward gDNA. 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. We demonstrate 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 gD
Czech name
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Czech description
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Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
EB - Genetics and molecular biology
OECD FORD branch
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Publication year
2012
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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