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' ¡æ 5' RNase H activity that degrades the RNA portion of
RNA-DNA hybrids.
SOURCE Purified from Avian Myeloblastosis virus.
APPLICATIONS ¡Ü cDNA synthesis. ¡Ü DNA
sequencing by dideoxy method. ¡Ü RNA sequencing. ¡Ü 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¡É.
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 ¥ìg/ml rA400:dT50.
CONCENTRATION: 5-10 units/¥ìl, 20-40 units/¥ìl (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¡É.¡¡
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.
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