Journal of Food, Agriculture and Environment
Vol 10,
Issue 3&4,2012
Online ISSN: 1459-0263
Print ISSN: 1459-0255
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A rapid DNA extraction method for quantitative real-time PCR amplification from fresh water sediment
Author(s):
Shimin Lu,
Mingjun Liao, Min Zhang, Pengzhi Qi, Congxin Xie*, Xugang He*Recieved Date: 2012-06-30, Accepted Date: 2012-09-30
Abstract:Many soil and sediment DNA extraction methods have been published and most of these methods evaluated the DNA purity by general PCR amplification, restriction enzyme reaction, or the ratio of A260/A280. However, these evaluation methods could not completely reflect the purity of sediment DNA to some extent, because of the existence of humic acid in the sample. Even trace levels of polluting chemicals in sediment can act as an inhibitor in the process of PCR. In this study, KAl(SO4)2 combined PEG6000 was used in sediment DNA extraction. The size of acquired DNA was determined by agarose gel electrophoresis; the yield and quality of DNA was measured with a UV-Vis spectrophotometer; and the purity of DNA was tested by quantitative real-time PCR (qPCR) of ammonia-oxidizing archaea (AOA) function gene amoA. Our method acquired a single and bright DNA band in 23 kb of size. The results also indicated that the DNA yield and purity could match the efficiency of the commercial sediment DNA extraction kit, but our method is much less expensive than the commercial kit.
Keywords:DNA extraction,
qPCR, sediment, AOAJournal: Journal of Food, Agriculture and Environment
Year: 2012
Volume: 10
Issue: 3&4
Category: Environment
Pages: 1252-1255
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