HR: 1340h
AN: B33A-0850 [Abstracts]
TI: Optimization of DNA Extraction from Deep-sea Basalt
AU: * Wang, H
EM: wanghmei04@163.com
AF: Geomicrobiology group, Department of Biological Sciences, Marine Environmental Section,
University of Southern California, 3616 Trousdale Parkway, Los Angeles, CA 90007, United States
AU: * Wang, H
EM: wanghmei04@163.com
AF: School of Environmental Studies, China University of Geosciences, No. 388 Lumo Road,
Wuhan Hubei province, 430074, China
AU: Edwards, K J
EM: kje@usc.edu
AF: Geomicrobiology group, Department of Biological Sciences, Marine Environmental Section,
University of Southern California, 3616 Trousdale Parkway, Los Angeles, CA 90007, United States
AB:
Studies on the microorganisms that inhabit deep-sea basalt can provide information on this dark ecosystem,
which will contribution to our understanding of mass transformation and energy flow in the deep ocean. However,
molecular methods for use with metal- and clay-rich rock materials such as basalt have not been suitably
developed at present, yet are critically required in order to be able to fully evaluate the basalt biotope. For
example, inefficient DNA extraction might lead to loss of information about important components of this
community, and misinterpretation about the total community diversity and function. In order to investigate the
effects of sample pretreated method, particle size, different DNA extraction methods and cell density on extracted
DNA yields, two basalt samples were collected from the East Pacific Rise 9° N during research cruise AT11-
20 in Nov 2004. Basalt samples were crushed to different particle size, washed with ddH2O and 100%
ethanol respectively, and autoclaved. Marinobacter aquaeolei cultures with different cell densities were
inoculated into differently treated basalt samples. Pure culture and basalt samples without inoculation were used
as positive and negative control to evaluate the extracting efficiency. FastDNA„¥ spin for soil kit, GeneClean„¥ for
ancient DNA kit and UltraCleanTM soil DNA Kit are used for DNA extraction. Results showed that DNA yields
increased with culture density. FastDNA„¥ spin for soil kit gave the highest DNA yields, which is almost 10 times
more than that of UltraCleanTM soil DNA Kit. Ethanol washing and ddH2O washing did not make big
difference to DNA yields. Mineral composition and surface areas might also affect DNA yields.
DE: 0448 Geomicrobiology
SC: Biogeosciences [B]
MN: 2007 Fall Meeting