Variables influencing the efficiency and interpretation of reverse transcription quantitative PCR (RT-qPCR): An empirical study using Bacteriophage MS2

•Increasing total RNA and primer concentrations improved RT efficiency.•At high concentrations, random hexamers performed as well as gene-specific primers.•RNA standards corrected for residual systematic variability in RT efficiency.•Length of the RNA standard dramatically influenced quantification...

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Bibliographic Details
Published in:	Journal of virological methods Vol. 241; pp. 1 - 10
Main Authors:	Miranda, Jaclyn A., Steward, Grieg F.
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 01.03.2017
Subjects:	bacteriophages cDNA synthesis Data Accuracy DNA Primers DNA, Complementary - genetics enzymes Levivirus - genetics quantitative polymerase chain reaction Reproducibility of Results reverse transcriptase polymerase chain reaction Reverse Transcriptase Polymerase Chain Reaction - methods Reverse Transcriptase Polymerase Chain Reaction - standards Reverse Transcription RNA RNA, Viral - genetics RNA, Viral - metabolism RT-qPCR Sensitivity and Specificity Virus quantification RT RNA rRNA s.d mRNA cDNA AMV Virus quantification RT-qPCR Cq MMLV cDNA synthesis
ISSN:	0166-0934, 1879-0984, 1879-0984
Online Access:	Get full text
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