title: Analysis of variant gene family expression by quantitative PCR.
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.
authors.affiliation: Department of Microbiology and Immunology, Weill Cornell Medical College, New York, NY, USA.
authors.forename: Sohini
authors.initials: S
authors.lastname: Sanyal
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Templeton
authors.forename: Cristina K
authors.initials: CK
authors.lastname: Moreira
mesh_terms: D019295:Computational Biology / Q000379:methods; D004912:Erythrocytes / Q000469:parasitology; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D017125:Genes, Protozoan; D006801:Humans; D005810:Multigene Family; D010961:Plasmodium / Q000235:genetics*; D016053:RNA, Protozoan / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D016053:RNA, Protozoan