The DOI matches its Crossref record; 15 of the 15 references it cites were found in a registry or an index.
High‐Throughput Real‐Time Quantitative Reverse Transcription PCR
Abstract
Extensive detail on the application of the real‐time quantitative polymerase chain reaction (QPCR) for the analysis of gene expression is provided in this unit. The protocols are designed for high‐throughput, 384‐well‐format instruments, such as the Applied Biosystems 7900HT, but may be modified to suit any real‐time PCR instrument. QPCR primer and probe design and validation are discussed, and three relative quantitation methods are described: the standard curve method, the efficiency‐corrected ΔCt method, and the comparative cycle time, or ΔΔCt method. In addition, a method is provided for absolute quantification of RNA in unknown samples. RNA standards are subjected to RT‐PCR in the same manner as the experimental samples, thus accounting for the reaction efficiencies of both procedures. This protocol describes the production and quantitation of synthetic RNA molecules for real‐time and non‐real‐time RT‐PCR applications.
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Cites
Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method
Livak, K. J., & Schmittgen, T. D. (2001) · Methods · doi:10.1006/meth.2001.1262 · Cited by 180,860
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[21] Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction
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Molecular Beacons: Probes that Fluoresce upon Hybridization
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Schmittgen, T. D., & Zakrajsek, B. A. (2000) · Journal of Biochemical and Biophysical Methods · doi:10.1016/s0165-022x(00)00129-9 · Cited by 1,242
Continuous Fluorescence Monitoring of Rapid Cycle DNA Amplification
Wittwer, C. T., Herrmann, M. G., Moss, A. A., & Rasmussen, R. P. (1997) · BioTechniques · doi:10.2144/97221bi01 · Cited by 1,143
Validation of housekeeping genes for normalizing RNA expression in real-time PCR
Dheda, K., Huggett, J. F., Bustin, S. A., Johnson, M. A., Rook, G., & Zumla, A. (2004) · BioTechniques · doi:10.2144/04371rr03 · Cited by 1,034
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Control Selection for RNA Quantitation
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Cited by
Anatomical profiling of nuclear receptor expression reveals a hierarchical transcriptional network
Bookout, A. L., Jeong, Y., Downes, M., Yu, R. T., Evans, R. M., & Mangelsdorf, D. J. (2006) · Cell · doi:10.1016/j.cell.2006.06.049 · Cited by 1,015
Self-citationRamirez-Farias, C., Slezak, K., Fuller, Z., Duncan, A., Holtrop, G., & Louis, P. (2008) · British Journal of Nutrition · doi:10.1017/s0007114508019880 · Cited by 822
Fon Tacer, K., Bookout, A. L., Ding, X., Kurosu, H., John, G. B., Wang, L., Goetz, R., Mohammadi, M., Kuro-o, M., Mangelsdorf, D. J., et al. (2010) · Molecular Endocrinology · doi:10.1210/me.2010-0142 · Cited by 700
Self-citationFGF21 regulates metabolism and circadian behavior by acting on the nervous system
Bookout, A. L., de Groot, M. H. M., Owen, B. M., Lee, S., Gautron, L., Lawrence, H. L., Ding, X., Elmquist, J. K., Takahashi, J. S., Mangelsdorf, D. J., et al. (2013) · Nature Medicine · doi:10.1038/nm.3249 · Cited by 545
Self-citationFGF21 acts centrally to induce sympathetic nerve activity, energy expenditure, and weight loss
Owen, B. M., Ding, X., Morgan, D. A., Coate, K. C., Bookout, A. L., Rahmouni, K., Kliewer, S. A., & Mangelsdorf, D. J. (2014) · Cell Metabolism · doi:10.1016/j.cmet.2014.07.012 · Cited by 523
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Metadata
- Authors
- Bookout, A. L., Cummins, C. L., Mangelsdorf, D. J., Pesola, J. M., & Kramer, M. F.
- Venue
- Current Protocols in Molecular Biology
- Publisher
- Wiley
- Published
- 2006-01
- Volume
- 73
- Issue
- 1
- Pages
- Unit 15.8-Unit 15.8
- ISSN
- 1934-3639 (ISSN-L)
- 1934-3647
- OpenAlex
- W2140734331
- Cited by
- 398
- References
- 15
- FWCI
- 4.05
- Type
- journal article
- Language
- en
- Open access
- closed
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