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RECOVERY OF DNA FROM SOILS AND SEDIMENTS
Citation:
Steffan, R., J. Goksoyr, A. Bej, AND R. Atlas. RECOVERY OF DNA FROM SOILS AND SEDIMENTS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-93/292 (NTIS PB93222834), 1988.
Description:
Experiments were performed to evaluate the effectiveness of different methodological approaches for recovering DNA from soil and sediment bacterial communities; cell extraction followed by lysis and DNA recovery (cell extraction method) versus direct cell lysis and alkaline extraction to recover DNA (direct lysis method). fficiency of DNA recovery by each method was determined by spectrophotometric absorbance using a tritiated thymidine tracer. ith both procedures, the use of polyvinylpolypyrrolidone was important for the removal of humic compounds to improve the purity of the recovered DNA; without extensive purification, various restriction enzymes failed to cut added target DNA. illigram quantities of high-purity DNA were recovered from 100-g samples of both soils and sediments by the direct lysis methods, which was a >1-order-of-magnitude-higher yield than by the cell extraction methods. he ratio of labeled thymidine to total DNA, however, was higher in the DNA recovered by the cell extraction method than by the direct lysis method, suggesting that the DNA recovered by the cell extraction method came primarily from active bacterial cells, whereas that recovered by the direct lysis method may have contained DNA from other sources.
