Fluorometric assay for DNA polymerases and reverse transcriptase.
Biotechniques
; 21(4): 664, 666, 668, 670, 672, 1996 Oct.
Article
in En
| MEDLINE
| ID: mdl-8891218
ABSTRACT
We report a quick, easy and inexpensive fluorometric assay that measures the activity of replication enzymes using Pico-GreenTM. The systems tested include replication of the natural template M13 Gori by E. coli DNA polymerase III holoenzyme and the replication of a synthetic homopolymer by human immunodeficiency virus reverse transcriptase. A direct comparison of the fluorometric assay with the conventional isotopic assay shows that the fluorometric assay accurately reflects the extent of replication. By performing the assay reactions directly in 96-well plates and using a fluorescence plate reader to determine the extent of reaction, the time required to measure replication activities is significantly shortened.
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Database:
MEDLINE
Main subject:
RNA-Directed DNA Polymerase
/
DNA-Directed DNA Polymerase
Limits:
Humans
Language:
En
Year:
1996
Type:
Article