Molecular Biotechnology

, Volume 8, Issue 1, pp 61–77

Reverse Transcriptase

The use of cloned moloney murine leukemia virus reverse transcriptase to synthesize DNA from RNA

Authors

    • Life Technologies
  • Donna K. Fox
    • Life Technologies
  • Margret Nathan
    • Life Technologies
  • James M. D'Alessio
    • Life Technologies
Protocol

DOI: 10.1007/BF02762340

Cite this article as:
Gerard, G.F., Fox, D.K., Nathan, M. et al. Mol Biotechnol (1997) 8: 61. doi:10.1007/BF02762340

Abstract

Reverse transcriptase (RT) is the key enzyme required for conversion of RNA to DNA. Cloning of Moloney murine leukemia virus (MMLV) RT has enable engineering an RT that lacks endogenous RNase H activity. RT catalyzes cDNA synthesis more efficiently in the absence of RNase H. We describe here a number of properties of MMLV RT and RNase H-minus MMLV RT not summarized in a single location elsewhere, providing a basis for best use of these enzymes in cDNA synthesis. In addition, general guidelines and detailed protocols are provided for use of MMLV RTs in one tube double-stranded cDNA synthesis, in [32P]cDNA synthesis, and in RT-PCR and long RT-PCR.

Index Entries

Murine leukemia virus reverse transcriptasereverse transcriptasecDNA synthesisRT-PCRlong RT-PCRreverse transcriptionRNase H-minus reverse transcriptase
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Copyright information

© Humana Press Inc. 1997