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Comparison of DNA extraction methods for the PCR-single-strand-conformation polymorphism analysis of kefir

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Abstract

Experiments were performed to determine the influence of three DNA extraction methods (i.e. lysozyme, sonication and CTAB methods) from kefir on the microbial diversity analysis by PCR-single strand conformation polymorphism (PCR-SSCP). The results showed that the band of DNA extracted using CTAB was clearer than that using other methods. In addition, the yield and purity of DNA extracted using CTAB were the highest and reached, respectively, 915 μg/ml and 1.694.The results from the experiments indicated that the CTAB-based DNA extraction method was the most efficient method for DNA extraction from kefir. The heterogeneity of PCR products, amplified from community DNA with universal primers spanning the V3 region of 16S rRNA genes, was analysed by using SSCP. The results showed that the SSCP profile based on the sonication method gave the highest microbial diversity of kefir. One conclusion from these results was that the DNA extraction method was an important factor affecting the SSCP-based microbial diversity analysis of kefir.

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Acknowledgments

Support for this research by the Shanxi Province Science Foundation (No.2008012010-2) is gratefully acknowledged.

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Authors and Affiliations

  1. School of Life Science and Technology, Shanxi University, Taiyuan, China

    Shu-Jun Chen & Jian-Rong Han

  2. Institute of Biotechnology, Shanxi University, Taiyuan, China

    Qing Wang, Yu Wang & Zhi-Wei Li

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  1. Shu-Jun Chen
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  2. Qing Wang
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  3. Jian-Rong Han
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  4. Yu Wang
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  5. Zhi-Wei Li
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Correspondence to Jian-Rong Han.

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Chen, SJ., Wang, Q., Han, JR. et al. Comparison of DNA extraction methods for the PCR-single-strand-conformation polymorphism analysis of kefir. World J Microbiol Biotechnol 27, 193–196 (2011). https://doi.org/10.1007/s11274-010-0438-z

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  • DOI: https://doi.org/10.1007/s11274-010-0438-z

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