High temperature cDNA synthesis by AMV reverse transcriptase improves the specificity of PCR.
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.
Fuchs
B
B
Department of Orthopedics, Mayo Clinic, Rochester, MN 55905, USA.
Zhang
K
K
Rock
M G
MG
Bolander
M E
ME
Sarkar
G
G
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
D018076:DNA, Complementary; D012194:RNA-Directed DNA Polymerase