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AMV Reverse Transcriptase, recombinant, GMP Grade

from Avian Myeloblastosis Virus, expressed in E. coli

For further processing only.

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Order information
AMV Reverse Transcriptase, recombinant, GMP Grade material number and pack size:
Material Number Pack Size
03203166103 20 kU or 200 kU
Will be supplied as ''Rev.Transcriptase, AMV rec.''. Unit of measure is ''kU''.
The enzyme is filled as 20 or 200 KU per vial. Specify in the order, which filling.
The enzyme is supplied without reaction buffer.
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Overview
AMV Reverse Transcriptase with high thermostability for generation of RNA fragments up to 12 kb with high sensitivity.
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Application
Use AMV Reverse Transcriptase for synthesis of cDNA from total RNA or mRNA for:
  • Two-step RT-PCR applications for amplification from RNA targets
  • RT-PCR for detection of viral RNA
  • TMA and NASBA nucleic acid amplification methods
  • Synthesis of full-length cDNA for libraries or cloning
  • Rapid amplification of cDNA end (RACE)
  • Manufacture of amplification mixtures for applications with regulatory requirements (e.g., in vitro diagnostics, quality control)
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Specification
AMV Reverse Transcriptase, recombinant, GMP Grade, provides higher thermostability compared to the native forms of AMV or M-MLV reverse transcriptase, allowing higher temperatures for reverse transcription, thus achieving better performance with GC-rich RNA fragments and difficult secondary structures.
Enzyme activities: RNA-dependent DNA polymerase, DNA-dependent DNA polymerase, unwinding activity, RNase H (degrading RNA in RNA:DNA hybrids)
Recommended reaction temperature: +42 to +65°C
Substrates: Incorporates dNTP, ddNTP, dUPT, various labeled or modified nucleotides
Divalent ion requirement: Mg2+
Appearance: Clear, colorless solution
Storage buffer: Potassium phosphate, 200 mmol/L; DTT, 2 mmol/L; Triton X-100, 0.2% (v/v); glycerol, 50% (v/v); pH approximately 7.2
Volume activity: ≥20 U/μL
Specific activity: ≥50 kU/mg protein
Unit definition: One unit AMV Reverse Transcriptase, recombinant, GMP Grade, is defined as the amount of enzyme which incorporates 1 nmol of [3H]TMP into an acid insoluble product in 10 minutes at +37°C with poly(A)x(dT)15 as substrate.
Purity (SDS PAGE): ≥90%
Unspecific endonucleases (MWM III DNA): Not detectable in up to 25 U after 16 hours incubation at +37°C.
Nicking activity (pBR322 DNA): Not detectable in up to 25 U after 16 hours incubation at +37°C.
Ribonucleases (MS2 RNA): Not detectable in up to 40 U after 4 hours incubation at +37°C.
Function test in RT-PCR (human skeletal muscle total RNA, 10 kb dystrophin gene fragment): Corresponds to reference
Bioburden: ≤50 CFU/mL
Animal-derived additives: None
Stability: At -15 to -25°C within specification range for 12 months.