Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays.

By Stephen A. Bustin. Journal of Molecular Endocrinology, 2000

4,175 citations recorded by the source corpus.

View Paper: https://www.semanticscholar.org/paper/c46e6f6bbfe2f0298cbc27e54f5ba7184d9cd018

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In Molecular Biology

Citations in the corpus

Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. (2002, methodology): “I have previously advocated normalisation to total cellular RNA as the least unreliable method (Bustin 2000).”
Conditional inactivation of FGF receptor 2 reveals an essential role for FGF signaling in the regulation of osteoblast function and bone growth (2003, methodology): “qRT-PCR was performed on an Applied Biosystems Gene Amp 5700 Sequence Detection system using Sybr Green fluorescent dye binding to PCR products (Bustin, 2000).”
Comparison of reference genes for quantitative real-time polymerase chain reaction analysis of gene expression in sugarcane (2007, methodology): “Ideally, a reference gene should be expressed at a relatively constant level in different tissues and at all stages of development (Bustin, 2000).”
Quantitative real-time RT-PCR--a perspective. (2005, methodology): “…model identifying GAPDH as one of the most suitable reference genes in colorectal cancer, when there is considerable evidence that mRNA levels of this gene vary signiﬁcantly between individuals and between paired normal and cancer tissue (Bustin et al. 1999, Bustin 2000, Tricarico et al. 2002).”
Experimental validation of novel and conventional approaches to quantitative real-time PCR data analysis. (2003, background): “The speed with which this technique has been adopted has led to a range of potential instrumentation with a corresponding variety of fluorescent chemistries ( 2 ,4).”
Real-time PCR in the microbiology laboratory. (2004, background): “Because the function of these oligoprobes depends upon correct hybridisation of the stem and two oligoprobes, their accurate design is crucial [8].”
Quantitative real-time reverse transcription polymerase chain reaction: normalization to rRNA or single housekeeping genes is inappropriate for human tissue biopsies. (2002, methodology): “We have previously shown that such short sense-strand amplicons, average length 70 bases, reliably mimic the RT step and generate standard curves that are identical to those obtained using T7-transcribed RNA [2].”
The power of real-time PCR. (2005, methodology): “This is called real-time RT-PCR and has become the most popular method of quantitating steady-state mRNA levels (9).”
Standard methods for molecular research in Apis mellifera (2013, methodology): “‘G’ bases should be avoided at the 5’ end, where the fluorophore usually resides, since they quench fluorescence, even after cleavage (Bustin, 2000).”
A new mathematical model for relative quantification in real-time RT-PCR. (2001, background): “Nevertheless, the generation of stable and reliable standard material, either recombinant DNA or recombinant RNA, is very time consuming and it must be precisely quantified (2, 7 ,8).”
A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything (2017, background): “This is due to its higher specificity mediated by the additional oligonucleotide— the probe—and the lower susceptibility to visualize non-specific PCR products, e.g., primer dimers (Bustin, 2000; Kubista et al., 2006).”
Comprehensive Algorithm for Quantitative Real-Time Polymerase Chain Reaction (2005, methodology): “Reagents have been developed ( Bustin, 2000, 2002 ) that can produce fluorescent signals proportional to the quantity of the amplicon over an appropriate range (Higuchi et al., 1993).”
Rapid in planta detection of Chalara fraxinea by a real-time PCR assay using a dual-labelled probe (2009, methodology): “... of several Chalara spp.) were aligned with additional ITS sequences obtained for several European C. fraxinea strains (see accessions in Table 1). A C. fraxinea-specific combination of dual labelled-probe and primer pair was manually designed (i) based upon species-specific polymorphisms reveale”
Pre-B cell colony-enhancing factor inhibits neutrophil apoptosis in experimental inflammation and clinical sepsis. (2004, methodology): “Real-time PCR was performed as described (65, 66) using SYBR Green PCR Master Mix (PerkinElmer Applied Biosystems, Warrington, United Kingdom) and”
Mapping of picoeucaryotes in marine ecosystems with quantitative PCR of the 18S rRNA gene. (2005, methodology): “In this work, we opted for the SYBR Green I approach [19], instead of the TaqMan approach, since the former requires only two primers for each target and therefore relies only on two conserved sites.”
How good is a PCR efficiency estimate: Recommendations for precise and robust qPCR efficiency assessments (2015, neutral): “E(E) = SE(slope) × (1 + E) ln 10 slope(2) (7)”