Quantitative real-time RT-PCR--a perspective.

By Stephen A. Bustin, Vladimir Benes, T. Nolan, M. W. Pfaffl. Journal of Molecular Endocrinology, 2005

1,367 citations recorded by the source corpus.

View Paper: https://www.semanticscholar.org/paper/f947a585fb5beb95eb78182d2a566c14a88f97c9

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Molecular Biology

Citations in the corpus

Reference genes in real-time PCR (2013, background): “It was also shown that use of random hexameric primer sequences for reverse transcription may overstate the actual amount of mRNA up to 19 times, therefore more accurate results might be obtained by using primers with sequences specific to a given area (Bustin et al. 2005).”
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, background): “At4g34270 and At4g33380 are two genes found to be stably expressed in Arabidopsis (Czechowski et al., 2005).”
Nucleic acid-based approaches to investigate microbial-related cheese quality defects (2012, background): “qPCR is rapid, extremely sensitive and has been applied in food microbiology, genomics, medicine, and environmental studies (Bustin et al., 2005; Quigley et al., 2011).”
PrimerBank: a resource of human and mouse PCR primer pairs for gene expression detection and quantification (2009, methodology): “Random priming can result in the highest coverage of the 5 0 -end and this method was used for our cDNA preparations (3,30).”
Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR (2006, background): “An accurate and relevant normalization to some internal standard is obligatory for biologically meaningful mRNA quantification (Bustin et al. 2005).”
Quantification of mRNA using real-time RT-PCR (2006, background): “and has serious implications for the reliability, relevance and reproducibility of RT-qPC”
RNA integrity and the effect on the real-time qRT-PCR performance. (2006, background): “The quality and quality of purified RNA is variable and after the extraction during long storage rather unstable (Bustin et al., 2005).”
Statistical analysis of real-time PCR data (2006, positive): “Recent advancements in real-time PCR primer design have allowed easier experimental optimization [16,17].”
Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon (2005, background): “This might have to do with the fact that geNorm will top-rank co-expressed genes [22], a weakness that has to be considered when evaluating paralog genes likely to be co-regulated.”
The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. (2009, background): “Consequently, although qPCR has frequently been proclaimed a touchstone or agoldstandard,inpracticethis“standard”isavariable one, and the reporting of results requires considerable sophistication of analysis and interpretation ( 43 ).”
Identification of suitable reference genes for studying gene expression in cucumber plants subjected to abiotic stress and growth regulators (2011, background): “The main advantages of this technique are high sensitivity, high specificity and broad quantification range (Bustin et al. 2005; Bustin and Nolan 2004; Gahon et al. 2004; Ginzinger 2002).”
An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development (2006, positive): “It provides outstanding accuracy of RNA quantification and has a broad dynamic range over wide experimental conditions [1-7].”
Molecular Techniques to Assess Microbial Community Structure, Function, and Dynamics in the Environment (2011, methodology): “markers (Bustin et al. 2005; Smith and Osborn 2009).”