RNA integrity and the effect on the real-time qRT-PCR performance.

By S. Fleige, M. W. Pfaffl. Molecular Aspects of Medicine, 2006

1,558 citations recorded by the source corpus.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/b66c425598171b4933686edbea8d918157d8fcd1

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

1. J. Huggett
2. T. Nolan
3. M. W. Pfaffl
4. S. Fleige
5. R. Hands
6. Jo Vandesompele
7. Jan Hellemans
8. Stephen A. Bustin

Citations in the corpus

• The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. (2009, background): “Instead, one should provide gel electrophoresis evidence at the least or, better yet, resultsfromamicrofluidics-basedrRNAanalysis( 50 )or a reference gene/target gene 3:5 integrity assay (51).”
• An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals. (2011, background): “The first issue is that RNA is easily degraded, and it is essential to ensure that sample collection strictly adheres to protocols designed for RNA preservation; this is particularly important when the samples are intended to be used for microarray analysis (47, 48).”
• Improving RNA-Seq expression estimates by correcting for fragment bias (2011, background): “Quantitative reverse transcription PCR (qRT-PCR) has served as a benchmark in numerous studies but it is not a perfect expression measurement assay [15], and it is therefore a priori unclear which technology currently produces the most accurate expression estimates.”
• Comparison of relative mRNA quantification models and the impact of RNA integrity in quantitative real-time RT-PCR (2006, background): “In cell culture and WBC the RIN ranged between 8.4 and 9.6 with low experimental variance (Fleige and Pfaffl 2006).”
• Quantification of mRNA using real-time RT-PCR (2006, methodology): “RNA assessment A variety of procedures are in common use for quantification of RN”
• Standard methods for molecular research in Apis mellifera (2013, background): “Such degradation can severely affect the accuracy and reliability of detecting and quantifying individual RNAs (Bustin and Nolan, 2004; Fleige and Pfaffl, 2006; Becker et al., 2010).”
• A practical approach to RT-qPCR-Publishing data that conform to the MIQE guidelines. (2010, positive): “Using partially degraded RNA can lead to varying and incorrect quantification results (Fleige and Pfaffl 2006, Gingrich et al. 2008).”