Quantification of mRNA using real-time RT-PCR

By T. Nolan, R. Hands, Stephen A. Bustin. Nature Protocols, 2006

2,269 citations recorded by the source corpus.

View Paper: https://www.semanticscholar.org/paper/be2adb9e4efdf1adbcf3fa0b9c088ac445c0d186

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Citations in the corpus

Touchdown PCR for increased specificity and sensitivity in PCR amplification (2008, background): “The only inherent limitation of the technique is that it is not suitable for quantitative assessment of target concentration, which is ideally performed by real-time PCR approaches under optimized condition”
A Transcription Activator-Like Effector (TALE) Toolbox for Genome Engineering (2012, methodology): “Although these assays are standard and have already been described elsewher”
Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data (2009, methodology): “During the last decade, quantitative real-time reverse transcriptase PCR, or qPCR for short, has become the method of choice for the quantification of mRNA transcripts (1,2).”
The lack of a systematic validation of reference genes: a serious pitfall undervalued in reverse transcription-polymerase chain reaction (RT-PCR) analysis in plants. (2008, methodology): “The development of realtime technologies, allowing quantitative RT-PCR, has provided complementary tools to microarrays, with consequent improvements in both sensitivity and specificity (Nolan et al ., 2006).”
qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data (2007, methodology): “Our quantification framework and software fit exactly in current thinking that places emphasis on getting every step of a realtime PCR assay right (such as RNA quality assessment, appropriate reverse transcription, selection of a proper normalization strategy, and so on [2]), especially if small dif”
The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. (2009, background): “Use of a 3:5 assay requires that the PCR efficiencies of both assays be virtually identical ( 51 ) and not be subject to differential inhibition.”
Comprehending crystalline β-carotene accumulation by comparing engineered cell models and the natural carotenoid-rich system of citrus (2011, neutral): “In the absence of an alternative, the use of a 3’:5’ assay has been proposed for assessing mRNA integrity (Nolan et al., 2006b).”
MIQE précis: Practical implementation of minimum standard guidelines for fluorescence-based quantitative real-time PCR experiments (2010, background): “Although no perfect quality assessment methods exist, measurements such as those obtained from microfluidics-based systems or 3’:5’-type assays can serve as basic indicators of RNA integrity [2].”