AMV Reverse Transcriptase

#3001

#3001HC

 

500 units

#3001L

#3001HCL

 

2,500 units

DESCRIPTION
AMV (Avian Myeloblastosis Virus) Reverse Transcriptase is an RNA-dependent DNA polymerase. This enzyme catalyses the polymerization of DNA using single-stranded DNA or RNA as a template in the presence of a primer. It also has a 3' 5' RNase H activity that degrades the RNA portion of RNA-DNA hybrids.

SOURCE
Purified from Avian Myeloblastosis virus.

APPLICATIONS
cDNA synthesis.
DNA sequencing by dideoxy method.
RNA sequencing.
3'-end labeling of DNA fragments.

UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 1nmol of dTTP into acid-insoluble form in 10 minutes at 37.

STANDARD UNIT ASSAY CONDITIONS
50mM Tris-HCl (pH8.3), 40mM KCl, 7mM MgCl2, 1mM DTT, 0.1mg/ml BSA, 0.5mM radiolabeled dTTP, and 35 g/ml rA400:dT50.

CONCENTRATION: 5-10 units/l, 20-40 units/l (High concentration product)

5X REACTION BUFFER 250mM Tris-HCl (pH8.3), 250mM KCl, 50mM MgCl2, 2.5mM spermidine, 50mM DTT.

STORAGE CONDITIONS
0.1M KPO4 (pH7.2), 0.2% Triton X-100, 2mM DTT, and 50% glycerol. Store at -70.

REFERENCES
1. Okayama, H. et al. (1987) Methods Enzymol. 154, 3.
2. Pfeffer, D. and Mierendorf, R. (1987) Methods Enzymol. 152, 563.
3. Houts, G.E. et al. (1979) J. Virol. 29, 517.