| Literature DB >> 26545322 |
Bhaja K Padhi1, Manjeet Singh2, Nicholas Huang2, Guillaume Pelletier2.
Abstract
Genomic DNA (gDNA) contamination of RNA samples can lead to inaccurate measurement of gene expression by reverse transcription quantitative real-time PCR (RT-qPCR). We describe an easily adoptable PCR-based method where gDNA contamination in RNA samples is assessed by comparing the amplification of intronic and exonic sequences from a housekeeping gene. Although this alternative assay was developed for rat RNA samples, it could be easily adapted to other species. As a proof of concept, we assessed the effects of detectable gDNA contamination levels on the expression of a few genes that illustrate the importance of RNA quality in acquiring reliable data. CrownEntities:
Keywords: Gene expression; Genomic DNA contamination; Pseudogene; RNA quality; Rat; Sdha
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Year: 2015 PMID: 26545322 DOI: 10.1016/j.ab.2015.10.012
Source DB: PubMed Journal: Anal Biochem ISSN: 0003-2697 Impact factor: 3.365