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AMV Reverse Transcriptase Online Order

AMV Reverse Transcriptase

AMV Reverse Transcriptase is isolated from the Avian Myeloblatosis Virus as the ab holoenzyme with a molecular weight of 157,000 daltons. AMV Reverse Transcriptase has both a 5’ 3’ primer-dependent polymerase activity which uses either RNA or DNA as a template, and a 3’ 5’ RNase H activity that degrades the RNA from RNA/DNA hybrids. It requires a primer as well as Mg2+ or Mn2+ ions. GENAXIS™ Biotechnology AMV Reverse Transcriptase can be applied to dideoxy sequencing of DNA and RNA and to cDNA synthesis using smaller RNA templates. In cDNA reactions, the enzyme is compatible with 4mM sodium pyrophosphate and placental RNase inhibitor. The optimal temperature is 41-45C. A temperature of 37-41C is preferable if NaPPi is used, however. GENAXIS™ Biotechnology AMV Reverse Transcriptase is delivered with a 10X reaction buffer.

APPLICATIONS: 

  • cDNA synthesisl
  • reverse transcription (RT-PCR)l
  • dideoxy sequencing of DNAl
  • primer extension and RNA sequencingl
  • 3’-end labeling of DNA fragments
CONTENTRATION:

    20 unit/l.

REACTION BUFFER 10X :

250mM Tris-HCl pH 8.3, 50mM MgCl2 , 500mM KCl, 20mM DTT.   

     

UNIT DEFINITION:

one unit is defined as the amount of enzyme required to incorporate 1 nmole dTTP into acid precipitable form after 10 minutes at 37C using poly(rA).p(dT)12-18 as template-primer.

COMMENTS:

purified free of endonucleases ; for first strand cDNA synthesis the following conditions are recommended : 50mM TrisHCl pH8.3, 8mM MgCl2, 30mM KCl, 10mM DTT, 2mM each dNTP and 20 units of enzyme per g of mRNA (incubate 1 hour at 42C).

STORAGE BUFFER:

    10mM KPO4 pH7.2, 2.0mM DTT, 2.0% (v/v) Triton X-100, 10% (v/v) glycerol.

STORAGE AND SHIPPING:

    store at –20C. Shipped refrigerated.

* PCR is a technology covered by patents owned by Hoffmann-La Roche Inc. and F. Hoffmann-La Roche Ltd.

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