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AMV Reverse Transcriptase, rec., concentrate

from Avian Myeloblastosis Virus, expressed in E. coli

For further processing only.

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Order information
AMV Reverse Transcriptase, rec., concentrate material number and pack size:
Material Number Pack Size
04460472103 200 kU
06676367103 20 kU
04460472103: Will be supplied as "AMV RT, rec., conc., 200 kU". Unit of measure is "kU".
06676367103: Will be supplied as "AMV RT, rec., conc., 20 kU". Unit of measure is "kU".
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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 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: ≥80-150 U/μL
Specific activity: ≥50 kU/mg protein
Unit definition: One unit AMV Reverse Transcriptase, recombinant, 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.