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A Hybrid DNA Extraction Method for the Qualitative and Quantitative Assessment of Bacterial Communities from Poultry Production Samples
Published on: December 10, 2014
Quantitative and qualitative validations of a sonication-based DNA extraction approach for PCR-based molecular
Xiaohu Dai1, Sisi Chen1, Ning Li1
1State Key Laboratory of Pollution Control and Resource Reuse, College of Environmental Science and Engineering, Tongji University, China.
Abstract:
The aim of this study was to comprehensively validate the sonication-based DNA extraction method, in hope of the replacement of the so-called 'standard DNA extraction method' - the commercial kit method. Microbial cells in the digested sludge sample, containing relatively high amount of PCR-inhibitory substances, such as humic acid and protein, were applied as the experimental alternatives. The procedure involving solid/liquid separation of sludge sample and dilution of both DNA templates and inhibitors, the minimum templates for PCR-based analyses, and the in-depth understanding from the bias analysis by pyrosequencing technology were obtained and confirmed the availability of the sonication-based DNA extraction method.

