Reverse Transcriptase AMV 10109118001

Enzyme Commission (EC) Number:

2.7.7.49 ( BRENDA | IUBMB )

Reverse Transcriptase, AMV is a gene product of the RNA genome of avian myeloblastosis virus. The enzymatically active forms of the purified enzyme are , and . The molecular weight of the -subunit is 68 kDa, that of the -subunit 92 kDa. The mature form, the most active form of Reverse Transcriptase, AMV, includes a RNA-directed DNA polymerase, a DNA-dependent DNA polymerase, a RNase H, and an unwinding activity. Reverse Transcriptase, AMV is used for cDNA synthesis, for synthesis of first strand cDNA for use in subsequent amplification reactions and dideoxy DNA sequencing.
The enzyme can also be used for RNA sequencing, 3 end labeling of DNA fragments, and the generation of ss probes for genomic footprints.
Reverse Transcriptase AMV requires a primer and Mg2+ or Mn2+ for activity.
Specificity
AMV (Avian Myeloblastosis Virus) Reverse Transcriptase is an RNA-directed DNA polymerase and a DNA-dependent DNA polymerase. The AMV reverse transcriptase requires a primer and Mg2+ or Mn2+ for activity.
Heat inactivation: 5 min, 95 °C
Reverse Transcriptase AMV is suitable for:

• First- and second-strand cDNA synthesis and synthesis of first strand cDNA for use in subsequent amplification reactions (RT-PCR)
• Dideoxy DNA sequencing
• Primer extension
• RNA sequencing
• 3-end labeling of DNA fragments
• Generation of single-stranded probes for genomic footprint experiments

Features and Benefits

• Efficiently transcribes total RNA, mRNA, viral RNA and RNA rich in secondary structures
• Procure full length cDNA fragments up to 12 kb
• Higher thermostability (up to 60°C) and specificity than M-MuLV Reverse Transcriptase

Packaging
1 kit containing 2 components
Quality
Absence of contaminants: Tested for the absence of detectable nonspecific RNases and nonspecific DNases in incubations with various nucleic acids (analyzed by gel electrophoresis) according to the current Quality Control procedures.
Function test: Reverse Transcriptase AMV is function tested in the cDNA Synthesis Kit and RT-PCR.
Unit Definition
1 unit is the enzyme activity that incorporates 1 nmol of [3H]-dTMP into acid-precipitable products in 10 minutes at +37 °C with poly(A)+ · d(pT)15 as template primer.
Volume Activity: 20-25 U/µl. Please refer to the Certificate of Analysis for more information.
Preparation Note
Storage buffer: 200 mM potassium phosphate, 2 mM dithiothreitol, 0.2% Triton X-100 (v/v), 50% glycerol (v/v), pH 7.2.
Storage and Stability
Store at -15–-25 °C. (Storage at -70 °C is more likely to deteriorate the enzyme via repeated freezing and thawing.)
For life science research only. Not for use in diagnostic procedures.

form	solution
Quality Level	100,
specific activity	>50 units/μg protein
feature	dNTPs included: no
packaging	pkg of 1,000 U
Торговая марка	Roche
storage condition	avoid repeated freeze/thaw cycles
parameter	42 °C optimum reaction temp.
application(s)	RT-qPCR: suitable
input	purified RNA
detection method	probe-based
shipped in	dry ice

RIDADR	NONH for all modes of transport
WGK Germany	WGK 1
Flash Point F	does not flash
Flash Point C	does not flash