Journal of Tropical Oceanography >
Evaluation of different extraction buffers on the quality of mangrove soil DNA
Received date: 2010-03-25
Revised date: 2010-05-19
Online published: 2011-05-04
Supported by
国家高技术研究发展计划项目(2008AA09Z408);国家自然科学基金项目(40930847、30930017、30940002); 深港创新圈项目(08Lh-04); 南方医科大学公共卫生与热带医学学院院长基金项目(GW200801-039)
The authors compared the influence of six different extraction buffers on DNA yields, DNA purity, co-extracted humic compound yields, and their PCR (polymerase chain reaction) sensitivity from a typical acidic and high organic content soil type, mangrove soil. Results showed that the quality of the soil DNA obtained using the acidic SDS (sodium dodecyl sul-fate) buffer was poor. The DNA yields obtained using the PVP (polyvinylpyrrolidone) buffer were higher than the other buff-ers, whereas the co-extracted humic compound yields were significantly higher. PCR results showed that no expected bands were obtained even in 1:1000 dilutions of the DNA extracts obtained using the acidic SDS buffer or PVP buffer. On the other hand, although the PVPP (polyvinylpolypyrrolidone) buffer and the CTAB (cetyltrimethylammonium bromide) and PVPP buffer resulted in a lower DNA yield, the purity of the soil DNA was improved. In addition, the purity was improved as the concentration of PVPP in the buffer increased. Moreover, PCR amplification was successfully conducted in the 1:10 dilutions of the DNA extracts obtained using the CTAB and PVPP buffer and in the 1:100 dilutions of DNA extracts obtained using the 2% PVPP buffer, respectively. In conclusion, acidic SDS buffer and PVP buffer were not suitable for extracting DNA from the acidic and high organic content soil type; however, 2% PVPP buffer and CTAB and PVPP buffer improved the DNA quality.
Key words： microbial diversity; DNA extraction; DNA yield; humic compound; extraction buffer
JIANG Yun-xia ,ZHENG Tian-ling , . Evaluation of different extraction buffers on the quality of mangrove soil DNA[J]. Journal of Tropical Oceanography, 2011 , 30(2) : 87 -93 . DOI: 10.11978/j.issn.1009-5470.2011.02.087
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