UNIT 3.7 RNA-Dependent DNA Polymerases
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Molecular Biology
How to Cite
Tabor, S. 2001. RNA-Dependent DNA Polymerases. Current Protocols in Molecular Biology. 00:III:3.7:3.7.1–3.7.3.
- Published Online: 1 MAY 2001
- Published Print: OCT 1987
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Reverse transcriptases are derived from retroviruses which use them to make DNA copies of their RNA genomes. They are multifunctional enzymes, but they are mainly used as RNA-directed DNA polymerases. Specifically, deoxyoligonucleotides (either oligo(dT) polymers, a collection of random DNA sequences, or a specific sequence) are used as primersfor extension on RNA (usually messenger RNA) templates. The DNA synthesized from the RNA templa te is referred to as complementary DNA (cDNA). This unit describes appropriate reaction conditions along with applications such as synthesizing cDNA for insertion into bacterial cloning vectors, filling in and labeling the 3′termini of DNA fragments with 5' protruding ends, and its use for DNA sequencing in place of Klenow fragment.