Fluorometric assay for DNA polymerases and reverse transcriptase. NLM AIDSLINE Important note: Information in this article was accurate in 1997. The state of the art may have changed since the publication date.

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Fluorometric assay for DNA polymerases and reverse transcriptase.

Biotechniques. 1996 Oct;21(4):664, 666, 668, 670, 672. Unique Identifier : AIDSLINE MED/97046297
Seville M; West AB; Cull MG; McHenry CS; Enzyco Inc., Denver, CO 80206, USA.

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.
Keywords: *DNA Polymerases/METABOLISM *RNA-Directed DNA Polymerase/METABOLISMKWDdnapolymerases/metabolismKWDrna-directeddnapolymerase/metabolism
970430
M9741607

Copyright © 1997 - National Library of Medicine. Reproduced under license with the National Library of Medicine, Bethesda, MD.

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