title: High temperature cDNA synthesis by AMV reverse transcriptase improves the specificity of PCR.
abstract: The enzyme avian myeloblastosis virus reverse transcriptase (AMV-RT) is routinely used for cDNA synthesis, which is generally carried out at temperatures between 37 degrees C and 42 degrees C. We show that this enzyme can support cDNA synthesis, at temperatures as high as 70 degrees C. We have utilized this property of the AMV-RT to improve the specificity of polymerase chain reaction (PCR). Furthermore, this apparently thermophilic property of the enzyme, which is an important constituent of a mesophilic organism, raises intriguing questions regarding evolution of the enzyme structure.
authors.affiliation: Department of Orthopedics, Mayo Clinic, Rochester, MN 55905, USA.
authors.forename: B
authors.initials: B
authors.lastname: Fuchs
authors.forename: K
authors.initials: K
authors.lastname: Zhang
authors.forename: M G
authors.initials: MG
authors.lastname: Rock
authors.forename: M E
authors.initials: ME
authors.lastname: Bolander
authors.forename: G
authors.initials: G
authors.lastname: Sarkar
mesh_terms: D009189:Avian Myeloblastosis Virus / Q000201:enzymology*; D018076:DNA, Complementary / Q000138:chemical synthesis*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013696:Temperature; D013698:Templates, Genetic
chemical_list: D018076:DNA, Complementary; D012194:RNA-Directed DNA Polymerase