AMV Reverse Transcriptase

Source

Avian Myeloblastosis Virus

Description

• Contains both reverse transcriptase and RNase H activities
• Maintains RNase H activity necessary for cDNA synthesis and isothermal RNA amplification

Applications

• Can be used for preparing labeled hybridization probes
• Synthesizes cDNA from single-stranded RNA
• Ideal for use in RT-PCR, RAMP™, NASBA™, cDNA libraries and dideoxy-DNA sequencing (1, 2, 3)

Reagents Supplied

5X AMV Reverse Transcriptase Reaction Buffer

Unit Definition

One unit is the amount of enzyme required to incorporate 1.0 nmol of labeled dTTP into acid insoluble material in 10 minutes at 37°C (4)

Concentration

20 - 30 units/µl

Reaction Buffer

250 mM Tris OAc, pH 8.4
375 mM KOAc
25 mM MgOAc

Storage Buffer

0.2 M potassium phosphate (pH 7.2)
2 mM dithiothreitol
0.2% Triton X-100™
50% (v/v) glycerol

Assay Conditions

50 mM Tris HCl (pH 8.3)
10 mM MgCl₂
40 mM KCl
0.5 mM [³H] dTTP
0.2 mM poly(rA):(dT)50
Reaction volume of 100 µl

Storage Conditions

Store at -20°C (non-frost-free)
Product shipped on Dry Ice