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PubMed · 8891218

Fluorometric assay for DNA polymerases and reverse transcriptase.

M Seville·A B West·M G Cull·C S McHenry
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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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M Seville, A B West, M G Cull, C S McHenry. 1996. Fluorometric assay for DNA polymerases and reverse transcriptase.. https://doi.org/10.2144/96214st04

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