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Underestimation of DNA Synthesis by [3H]Thymidine Incorporation in Marine Bacteria
Journal article   Peer reviewed

Underestimation of DNA Synthesis by [3H]Thymidine Incorporation in Marine Bacteria

Wade H. Jeffrey and John H. Paul
Applied and environmental microbiology, Vol.54(12), pp.3165-3168
12/01/1988
DOI: https://doi.org/10.1128/AEM.54.12.3165-3168.1988
PMCID: PMC204444
PMID: 16347806
Web of Science ID: WOS:A1988R141800046

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Abstract

A direct comparison of [3H]thymidine incorporation with DNA synthesis was made by using an exponentially growing estuarine bacterial isolate and the naturally occurring bacterial populations in a eutrophic subtropical estuary and in oligotrophic offshore waters. Simultaneous measurements of [3H]thymidine incorporation into DNA, fluorometrically determined DNA content, and direct counts were made over time. DNA synthesis estimated from thymidine incorporation values was compared with fluorometrically determined changes in DNA content. Even after isotope dilution, nonspecific macromolecular labeling, and efficiency of DNA recovery were accounted for, [3H]thymidine incorporation consistently underestimated DNA synthesized by six- to eightfold. These results indicate that although the relationship of [3H]thymidine incorporation to DNA synthesis appears consistent, there are significant sources of thymine bases incorporated into DNA which cannot be accounted for by standard [3H]thymidine incorporation and isotope dilution assays.

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