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M-MLV Reverse Transcriptase, GMP Grade

from Moloney Murine Leukemia Virus, expressed in E. coli

For further processing only.

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Order information
M-MLV Reverse Transcriptase, GMP Grade material number and pack size:
Material Number Pack Size
04707486103 200 kU
Will be supplied as "M-MLV RT Industrial GMP Grade, 200 KU". Unit of measure is "piece".
The enzyme is supplied without reaction buffer.
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Overview
M-MLV Reverse Transcriptase for preparation of full-length cDNA with high efficiency.
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Application
Use M-MLV 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
M-MLV Reverse Transcriptase, GMP Grade, is highly processive and generates full length cDNA with high efficiency. It has a lower RNase H activity than AMV Reverse Transcriptase and lacks endonuclease activity.
Enzyme activities: RNA-dependent DNA polymerase, DNA-dependent DNA polymerase, low RNase H activity, no endonuclease activity
Recommended reaction temperature: +37°C
Substrates: Incorporates dNTP, ddNTP, dUPT, various labeled or modified nucleotides
Divalent ion requirement: Mg2+
Appearance: Clear, colorless solution
Storage buffer: Tris/HCl, 25 mmol/L; NaCl, 100 mmol/L; DTT, 10 mmol/L; EDTA, 0.1 mmol/L; Triton X-100, 0.01% (v/v); glycerol, 50% (v/v); pH approximately 7.5
Volume activity: 200-300 U/μL
Specific activity: ≥100 kU/mg protein
Unit definition: One unit M-MLV Reverse Transcriptase, 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 100 U after 16 hours incubation at +37°C.
Nicking activity (pBR322 DNA): Not detectable in up to 100 U after 16 hours incubation at +37°C.
Ribonucleases (MS2 RNA): Not detectable in up to 200 U after 1 hour incubation at +37°C.
Bioburden: ≤50 CFU/mL
Animal-derived additives: None
Stability: At -15 to -25°C within specification range for 12 months.