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Properties of Reverse Transcriptases

Enzyme Maximum size of template Temperature optimum Units needed for standard reaction Ion require-
ment
Rnase H activity Combined RT and DNA Pol. Incorporates DIG-11-dUTP Expand Re-
verse Trans-
criptased up to 14kb 42C (up to 50C) 50 Mg2+ - - yes AMV Reverse Transcriptase up to 12kb 42C (up to 60C) 40 Mg2+ ++ - yes M-MuLV Reverse Transcriptase up to 10kb 37C 40 Mg2+ + - yes C. therm Polymerase for RT in Two-Step RT-PCR up to 4kb 60-70C 6 Mg2+ - -e yes Tth DNA Polymerase up to 1kb 60-70C 4 Mn2+ - + yes a Number in parentheses indicates maximum temperature tolerated by enzyme. b Standard reaction performed as recommended in enzyme package insert. c For nonradioact ive labeling; also incorporates other modified nucleotides. d Not for sale in the U.S. e C. therm. Polymerase for Reverse Transcription in Two-Step RT-PCR does not survive incubation at 95C. For RT-PCR, use the polymerase together with a second, thermostable enzyme such as Taq DNA Polymerase or Expand High Fidelity PCR System.
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2. Reverse transcriptases suitable for one-step RT-PCRb
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