【摘要】 分别采用两种环境总DNA直接提取法和一种间接提取法从6种温泉菌席样品中提取总DNA,以DNA粗产物的纯度、能否用于后续PCR扩增及PCR-DGGE(变性梯度凝胶电泳)所反映的微生物多样性为评价指标对两类方法进行比较和评价。研究发现,虽然间接提取法效率低下,但对于高温极端环境中生物量较小的样品,间接法能得到有研究价值的、纯度较高的环境样品总DNA,而直接法得到的DNA量小且不适于PCR扩增操作。在使用这2类方法都能得到可用于研究操作的DNA的情况下,间接提取法能更好的体现环境样品中微生物的多样性。更多还原

【Abstract】 Two different direct DNA extraction methods and one indirect DNA extraction method were applied to recover environmental DNA from six mat samples, which were sampled closely around a high-temperature spring vent in Tengchong Rehai of the western Yunnan province. As the criteria, quantity and purity of the extracted crude DNA, the result of PCR amplification for denaturing gradient gel electrophoresis (DGGE) before and after crude DNA purification, and the Shannon-wiener index of DGGE profiles were used to evaluate the direct and indirect DNA extraction methods. For the samples of less biomass, the indirect method yielded available crude DNA with high purity, and the DNA was amplified without purification by PCR for DGGE. The two direct methods extracted less DNA than indirect method from the samples of less biomass, and the crude DNA extraction could not be amplified by PCR. Despite the lower quantity of DNA yield, the indirect DNA extraction method for the other mat samples presented more diverse bacterial community than that of the direct DNA extraction methods according to the Shannon-wiener index of the PCR-DGGE profiles.更多还原