[Comparison of direct and indirect environmental DNA extraction methods for samples from a high-temperature environment].
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.
Liu
Hong-wei
HW
Yunnan Institute of Microbiology, Yunnan University, Kunming 650091, China. cliff.liu@163.com
Peng
Qian
Q
Li
Qin-yuan
QY
Xiao
Wei
W
Cui
Xiao-long
XL
Lin
Lian-bing
LB
Liu
Zhi-jian
ZJ
Jiang
Cheng-lin
CL
Xu
Li-hua
LH
Chen
Yi-guang
YG
Wang
Zhi-gang
ZG
Ren
Zhen
Z
Deng
Lan
L
Wen
Meng-liang
ML
D004269:DNA, Bacterial / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D004783:Environmental Microbiology; D045482:Hot Springs / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D012988:Soil Microbiology
D004269:DNA, Bacterial