UNIT 3.7 RNA-Dependent DNA Polymerases
Published Online: 1 OCT 2008
Copyright © 2008 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Tzertzinis, G., Tabor, S. and Nichols, N. M. 2008. RNA-Dependent DNA Polymerases. Current Protocols in Molecular Biology. 84:3.7.1–3.7.4.
- Published Online: 1 OCT 2008
- Published Print: OCT 2008
Reverse transcriptases (RTs) are multifunctional enzymes, but are mainly used as RNA-directed DNA polymerases in first-strand cDNA synthesis. Specifically, oligodeoxynucleotides are used as primers for extension on RNA templates. The DNA synthesized from an RNA template is referred to as complementary DNA (cDNA) and is often used as a template for PCR or converted to dsDNA for cloning. This unit describes appropriate reaction conditions for RTs from Moloney murine leukemia virus (MMLV) and avian myeloblastosis virus (AMV), along with applications such as synthesizing cDNA, 3′ fill-in reactions, and labeling the 3′ terminus of DNA fragments with 5′ protruding ends, and DNA sequencing. Curr. Protoc. Mol. Biol. 84:3.7.1-3.7.4. © 2008 by John Wiley & Sons, Inc.
- first-strand cDNA;
- MMLV RT;
- AMV RT;
- fill-in reaction