title: Reverse transcription-polymerase chain reaction.
abstract: Reverse transcription coupled to the polymerase chain reaction (RT-PCR) is commonly used to detect the presence of mRNAs, pre-mRNAs, or other types of RNA such as noncoding RNAs. The method involves using a primer annealed to the RNA of interest. For mRNA, the primer is usually a synthetic oligo(dT)15-18, a random hexamer mixture (dN)6, or a synthetic DNA oligonucleotide that is complementary to a specific transcript (a gene-specific primer). This DNA:RNA hybrid serves as a template during reverse transcription, in which the enzyme reverse transcriptase (RT) generates a single-stranded cDNA copy of a portion of the target RNA molecule. Using random hexamer priming, it is possible to obtain representative cDNA copies of sequences from the entire length of the mRNAs and pre-mRNAs in a population. This cDNA can then be used as a template for PCR. On addition of gene-specific primers, a specific DNA fragment corresponding to a portion of the RNA of interest is generated.
authors.forename: Donald C
authors.initials: DC
authors.lastname: Rio
mesh_terms: D017931:DNA Primers / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000032:analysis* / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D012313:RNA; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase