Recovery of DNA from Soils and Sediments

摘要

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). Efficiency of DNA recovery by each method was determined by spectrophotometric absorbance using a tritiated thymidine tracer. With 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. Milligram 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. (Copyright (c) 1988, American Society for Microbiology.)