Real-time PCR for mRNA quantitation - PubMed (original) (raw)

Review

doi: 10.2144/05391RV01.

Affiliations

• PMID: 16060372
• DOI: 10.2144/05391RV01

Free article

Review

Real-time PCR for mRNA quantitation

Marisa L Wong et al. Biotechniques. 2005 Jul.

Free article

Abstract

Real-time PCR has become one of the most widely used methods of gene quantitation because it has a large dynamic range, boasts tremendous sensitivity, can be highly sequence-specific, has little to no post-amplification processing, and is amenable to increasing sample throughput. However, optimal benefit from these advantages requires a clear understanding of the many options available for running a real-time PCR experiment. Starting with the theory behind real-time PCR, this review discusses the key components of a real-time PCR experiment, including one-step or two-step PCR, absolute versus relative quantitation, mathematical models available for relative quantitation and amplification efficiency calculations, types of normalization or data correction, and detection chemistries. In addition, the many causes of variation as well as methods to calculate intra- and inter-assay variation are addressed.

PubMed Disclaimer

Comment in

• Comment on Wong and Medrano's "Real-time PCR for mRNA quantification" BioTechniques 39:75-85 (July 2005).
  Dufva M. Dufva M. Biotechniques. 2005 Oct;39(4):484; discussion 484-5. doi: 10.2144/000112029. Biotechniques. 2005. PMID: 16235559 No abstract available.

Similar articles

• Comment on Wong and Medrano's "Real-time PCR for mRNA quantification" BioTechniques 39:75-85 (July 2005).
  Dufva M. Dufva M. Biotechniques. 2005 Oct;39(4):484; discussion 484-5. doi: 10.2144/000112029. Biotechniques. 2005. PMID: 16235559 No abstract available.
• Quantitative (real-time) RT-PCR in cardiovascular research.
  Ashton KJ, Headrick JP. Ashton KJ, et al. Methods Mol Biol. 2007;366:121-43. doi: 10.1007/978-1-59745-030-0_7. Methods Mol Biol. 2007. PMID: 17568122
• A standard curve based method for relative real time PCR data processing.
  Larionov A, Krause A, Miller W. Larionov A, et al. BMC Bioinformatics. 2005 Mar 21;6:62. doi: 10.1186/1471-2105-6-62. BMC Bioinformatics. 2005. PMID: 15780134 Free PMC article.
• The real-time polymerase chain reaction.
  Kubista M, Andrade JM, Bengtsson M, Forootan A, Jonák J, Lind K, Sindelka R, Sjöback R, Sjögreen B, Strömbom L, Ståhlberg A, Zoric N. Kubista M, et al. Mol Aspects Med. 2006 Apr-Jun;27(2-3):95-125. doi: 10.1016/j.mam.2005.12.007. Epub 2006 Feb 3. Mol Aspects Med. 2006. PMID: 16460794 Review.
• Complementary techniques: validation of gene expression data by quantitative real time PCR.
  Provenzano M, Mocellin S. Provenzano M, et al. Adv Exp Med Biol. 2007;593:66-73. doi: 10.1007/978-0-387-39978-2_7. Adv Exp Med Biol. 2007. PMID: 17265717 Review.

Cited by

• Impact of One-Week Administration of Dihydrotestosterone in Rat Anterior Pituitary Gland.
  Kanasaki H, Tumurbaatar T, Cairang Z, Tumurgan Z, Oride A, Okada H, Kyo S. Kanasaki H, et al. Int J Endocrinol. 2022 Oct 21;2022:9525227. doi: 10.1155/2022/9525227. eCollection 2022. Int J Endocrinol. 2022. PMID: 36311909 Free PMC article.
• Focused role of nanoparticles against COVID-19: Diagnosis and treatment.
  Dheyab MA, Khaniabadi PM, Aziz AA, Jameel MS, Mehrdel B, Oglat AA, Khaleel HA. Dheyab MA, et al. Photodiagnosis Photodyn Ther. 2021 Jun;34:102287. doi: 10.1016/j.pdpdt.2021.102287. Epub 2021 Apr 6. Photodiagnosis Photodyn Ther. 2021. PMID: 33836276 Free PMC article. Review.
• Pulchrin A, a New Natural Coumarin Derivative of Enicosanthellum pulchrum, Induces Apoptosis in Ovarian Cancer Cells via Intrinsic Pathway.
  Nordin N, Fadaeinasab M, Mohan S, Mohd Hashim N, Othman R, Karimian H, Iman V, Ramli N, Mohd Ali H, Abdul Majid N. Nordin N, et al. PLoS One. 2016 May 2;11(5):e0154023. doi: 10.1371/journal.pone.0154023. eCollection 2016. PLoS One. 2016. PMID: 27136097 Free PMC article.
• Evaluation and Selection of Appropriate Reference Genes for Real-Time Quantitative PCR Analysis of Gene Expression in Nile Tilapia (Oreochromis niloticus) during Vaccination and Infection.
  Wang E, Wang K, Chen D, Wang J, He Y, Long B, Yang L, Yang Q, Geng Y, Huang X, Ouyang P, Lai W. Wang E, et al. Int J Mol Sci. 2015 Apr 30;16(5):9998-10015. doi: 10.3390/ijms16059998. Int J Mol Sci. 2015. PMID: 25941937 Free PMC article.
• MeCP2 binds to nucleosome free (linker DNA) regions and to H3K9/H3K27 methylated nucleosomes in the brain.
  Thambirajah AA, Ng MK, Frehlick LJ, Li A, Serpa JJ, Petrotchenko EV, Silva-Moreno B, Missiaen KK, Borchers CH, Adam Hall J, Mackie R, Lutz F, Gowen BE, Hendzel M, Georgel PT, Ausió J. Thambirajah AA, et al. Nucleic Acids Res. 2012 Apr;40(7):2884-97. doi: 10.1093/nar/gkr1066. Epub 2011 Dec 5. Nucleic Acids Res. 2012. PMID: 22144686 Free PMC article.

Publication types

MeSH terms

Substances

LinkOut - more resources

• Full Text Sources

  • Atypon

• Other Literature Sources

  • The Lens - Patent Citations Database