Substantial performance discrepancies among commercially available kits for reverse transcription quantitative polymerase chain reaction: A systematic comparative investigator-driven approach

Analytical Biochemistry

Volumn 401, Issue 2, 2010, Pages 303-311

  Sieber, Matthias W ^a   Recknagel, Peter ^a   Glaser, Florian ^a   Witte, Otto W ^a   Bauer, Michael ^a   Claus, Ralf A ^a   Frahm, Christiane ^a  

^a JENA UNIVERSITY HOSPITAL   (Germany)

Author keywords

CDNA synthesis; Efficiency; Quantitative PCR; Real time PCR; Reproducibility; Reverse transcription; Rt qPCR; Sensitivity

Indexed keywords

EFFICIENCY; RNA;

CDNA SYNTHESIS; QUANTITATIVE PCR; REAL-TIME PCR; REPRODUCIBILITIES; REVERSE TRANSCRIPTION; RT-QPCR; SENSITIVITY;

POLYMERASE CHAIN REACTION;

MESSENGER RNA;

ACCURACY; ANALYTICAL EQUIPMENT; ANIMAL TISSUE; ARTICLE; CONTROLLED STUDY; HUMAN; HUMAN TISSUE; MOUSE; NONHUMAN; NUCLEOTIDE SEQUENCE; PRIORITY JOURNAL; PROCESS OPTIMIZATION; PRODUCTIVITY; RAT; RELIABILITY; REPRODUCIBILITY; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; SENSITIVITY ANALYSIS; TISSUE DISTRIBUTION; VALIDATION PROCESS;

RATTUS;

EID: 77952886657     PISSN: 00032697     EISSN: 10960309     Source Type: Journal    
DOI: 10.1016/j.ab.2010.03.007     Document Type: Article

References (27)

1. Bustin S.A., Benes V., Garson J.A., Hellemans J., Huggett J., Kubista M., Mueller R., Nolan T., Pfaffl M.W., Shipley G.L., Vandesompele J., Wittwer C.T. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin. Chem. 2009, 55:611-622.
2. Alwine J.C., Kemp D.J., Stark G.R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc. Natl. Acad. Sci. USA 1977, 74:5350-5354.
3. Gall J.G., Pardue M.L. Formation and detection of RNA-DNA hybrid molecules in cytological preparations. Proc. Natl. Acad. Sci. USA 1969, 63:378-383.
4. Pardue M.L., Gall J.G. Molecular hybridization of radioactive DNA to the DNA of cytological preparations. Proc. Natl. Acad. Sci. USA 1969, 64:600-604.
5. Berk A.J., Sharp P.A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell 1977, 12:721-732.
6. Schena M., Shalon D., Davis R.W., Brown P.O. Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science 1995, 270:467-470.
7. Velculescu V.E., Zhang L., Vogelstein B., Kinzler K.W. Serial analysis of gene expression, Science 1995, 270:484-487.
8. R. Higuchi, C. Fockler, G. Dollinger, R. Watson, Kinetic PCR analysis: real-time monitoring of DNA amplification reactions, Biotechnology (NY) 11 (1993) 1026-1030.
9. Heid C.A., Stevens J., Livak K.J., Williams P.M. Real time quantitative PCR. Genome Res. 1996, 6:986-994.
10. Bustin S.A. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J. Mol. Endocrinol. 2000, 25:169-193.
11. Streit S., Michalski C.W., Erkan M., Kleeff J., Friess H. Northern blot analysis for detection and quantification of RNA in pancreatic cancer cells and tissues. Nat. Protoc. 2009, 4:37-43.
12. Rottman J.B. The ribonuclease protection assay: a powerful tool for the veterinary pathologist. Vet. Pathol. 2002, 39:2-9.
13. Velculescu V.E., Vogelstein B., Kinzler K.W. Analysing uncharted transcriptomes with SAGE. Trends Genet. 2000, 16:423-425.
14. Ginzinger D.G. Gene quantification using real-time quantitative PCR: An emerging technology hits the mainstream. Exp. Hematol. 2002, 30:503-512.
15. Gibson U.E., Heid C.A., Williams P.M. A novel method for real time quantitative RT-PCR. Genome Res. 1996, 6:995-1001.
16. Hayward-Lester A., Oefner P.J., Sabatini S., Doris P.A. Accurate and absolute quantitative measurement of gene expression by single-tube RT-PCR and HPLC. Genome Res. 1995, 5:494-499.
17. Schroeder A., Mueller O., Stocker S., Salowsky R., Leiber M., Gassmann M., Lightfoot S., Menzel W., Granzow M., Ragg T. The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol. Biol. 2006, 7:3.
18. Pfaffl M.W., Feige S., Riedmaier I. Validation of lab-on-chip capillary electrophoresis systems for total RNA quality and quantity control. Biotechnol. Biotechnol. Equip. 2008, 22:829-834.
19. Stahlberg A., Hakansson J., Xian X., Semb H., Kubista M. Properties of the reverse transcription reaction in mRNA quantification. Clin. Chem. 2004, 50:509-515.
20. Rozen S., Skaletsky H. Primer3 on the WWW for general users and for biologist programmers. Methods Mol. Biol. 2000, 132:365-386.
21. Sieber M.W., Guenther M., Kohl M., Witte O.W., Claus R.A., Frahm C. Inter-age variability of bona fide unvaried transcripts: Normalization of quantitative PCR data in ischemic stroke. Neurobiol. Aging 2010, 31:654-664.
22. Pfaffl M.W. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001, 29:e45.
23. Kubista M., Andrade J.M., Bengtsson M., Forootan A., Jonak J., Lind K., Sindelka R., Sjoback R., Sjogreen B., Strombom L., Stahlberg A., Zoric N. The real-time polymerase chain reaction. Mol. Aspects Med. 2006, 27:95-125.
24. Gerard G.F., Fox D.K., Nathan M., D'Alessio J.M. Reverse transcriptase: the use of cloned Moloney murine leukemia virus reverse transcriptase to synthesize DNA from RNA. Mol. Biotechnol. 1997, 8:61-77.
25. Burell M.B. Enzymes of molecular biology. Methods Mol. Biol. 1994, 16:384.
26. Suslov O., Steindler D.A. PCR inhibition by reverse transcriptase leads to an overestimation of amplification efficiency. Nucleic Acids Res 2005, 33:e181.
27. Tichopad A., Kitchen R., Riedmaier I., Becker C., Stahlberg A., Kubista M. Design and optimization of reverse-transcription quantitative PCR experiments. Clin. Chem. 2009, 55:1816-1823.