Literature DB >> 26545322

A PCR-based approach to assess genomic DNA contamination in RNA: Application to rat RNA samples.

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. Crown
Copyright © 2015. Published by Elsevier Inc. All rights reserved.

Entities:  

Keywords:  Gene expression; Genomic DNA contamination; Pseudogene; RNA quality; Rat; Sdha

Mesh:

Substances:

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


  4 in total

1.  A PCR-based quantitative assay for the evaluation of mRNA integrity in rat samples.

Authors:  Bhaja K Padhi; Manjeet Singh; Marianela Rosales; Guillaume Pelletier; Sabit Cakmak
Journal:  Biomol Detect Quantif       Date:  2018-03-16

2.  Assessment of DNA Contamination in RNA Samples Based on Ribosomal DNA.

Authors:  Seyyed Hamidreza Hashemipetroudi; Ghorbanali Nematzadeh; Gholamreza Ahmadian; Ahad Yamchi; Markus Kuhlmann
Journal:  J Vis Exp       Date:  2018-01-22       Impact factor: 1.355

3.  MicroRNA miR-4709-3p targets Large Tumor Suppressor Kinase 2 (LATS2) and induces obstructive renal fibrosis through Hippo signaling.

Authors:  Zexiang Jiang; Weiping Xia; Guoyu Dai; Bo Zhang; Yang Li; Xiang Chen
Journal:  Bioengineered       Date:  2021-12       Impact factor: 3.269

4.  Genes expressed at low levels raise false discovery rates in RNA samples contaminated with genomic DNA.

Authors:  Xiangnan Li; Peipei Zhang; Haijian Wang; Ying Yu
Journal:  BMC Genomics       Date:  2022-08-03       Impact factor: 4.547

  4 in total

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