Abstract
Real-time polymerase chain reaction (PCR), or quantitative PCR (qPCR), is a rapid, sensitive, and specific method used for a broad variety of applications including quantitative gene expression analysis, DNA copy number measurement, characterization of gene and chromosomal deletions, and genotyping. Real-time reverse transcription (RT)-PCR has largely supplanted Northern blot and RNase protection assays, as two examples, as a means of quantifying transcript levels. The method utilizes small amounts of RNA and allows efficient screening of a large number of samples. Here, we describe the materials and methods required to perform real-time RT-PCR, including RNA purification, cDNA synthesis, and real-time PCR analysis of cDNA samples.
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Acknowledgments
This work was supported by NIH/NIAID grant 1R01AI080754-01A1 and the William Randolph Hearst Foundation to Thomas Templeton and Sohini Sanyal, and by NIH/NCRR CTSC grant UL1 RR 024996 to Cristina Moreira.
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Sanyal, S., Templeton, T.J., Moreira, C.K. (2012). Analysis of Variant Gene Family Expression by Quantitative PCR. In: Ménard, R. (eds) Malaria. Methods in Molecular Biology, vol 923. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-026-7_12
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DOI: https://doi.org/10.1007/978-1-62703-026-7_12
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-025-0
Online ISBN: 978-1-62703-026-7
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