UNIT 15.8 High-Throughput Real-Time Quantitative Reverse Transcription PCR
Published Online: 1 AUG 2005
Copyright © 2005 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Bookout, A. L., Cummins, C. L. and Mangelsdorf, D. J. 2005. High-Throughput Real-Time Quantitative Reverse Transcription PCR. Current Protocols in Molecular Biology. 15:15.8.
- Published Online: 1 AUG 2005
- Published Print: JUL 2005
This is not the most recent version of the article. View current version (1 FEB 2006)
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 ΔCt method.
- quantitative PCR;
- real-time PCR;
- RNA expression analysis;
- SYBR Green;
- ABI 7900HT;
- reverse transcriptase PCR