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PCR–DGGE method for analyzing the bacterial community in a high temperature petroleum reservoir

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Abstract

Different PCR–denaturing gradient gel electrophoresis (DGGE) protocols were employed to investigate bacterial communities in a high temperature and water flooded petroleum reservoir in Dagang oil field, China. Bacterial universal primers sets frequently used in PCR–DGGE were evaluated. Three primers sets P1 (341F-GC and 534R), P2 (341F-GC and 907R) and P3 (1055F and 1406R-GC) showed different DGGE patterns. Good separation and quality of patterns were obtained in DGGE analysis with the set P3. A total of 12 DNA fragments were excised from the DGGE gels and their sequences were determined. Clustering analysis of the DGGE profiles showed that bacteria in this petroleum reservoir belonged to four clusters. These results indicate that the procedure of DGGE analysis with the primer P3 (1055F and 1406R-GC) is suitable for investigating microbial community in petroleum reservoirs.

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Abbreviations

DGGE:

Denaturing gradient gel electrophoresis

CL:

Chemical lysis method

LN:

Liquid nitrogen lysis method

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Acknowledgements

This work was supported by grant from the National Natural Science Foundation (50674058) and the Tianjin Key Project of Applied Basic Research Effort, (07JCZDJC03200).

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Authors and Affiliations

  1. Tianjin Key Laboratory for Microbial Functional Genomics, Key Laboratory of Molecular Microbiology Technology, Ministry of Education, College of Life Sciences, Nankai University, 94, Weijin Road, Tianjin, 300071, China

    Jun Wang, Ting Ma, Lingxia Zhao, Jinghua Lv, Guoqiang Li, Fenglai Liang & Rulin Liu

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  1. Jun Wang
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  2. Ting Ma
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  3. Lingxia Zhao
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  4. Jinghua Lv
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  5. Guoqiang Li
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  6. Fenglai Liang
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  7. Rulin Liu
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Correspondence to Rulin Liu.

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Wang, J., Ma, T., Zhao, L. et al. PCR–DGGE method for analyzing the bacterial community in a high temperature petroleum reservoir. World J Microbiol Biotechnol 24, 1981–1987 (2008). https://doi.org/10.1007/s11274-008-9694-6

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  • DOI: https://doi.org/10.1007/s11274-008-9694-6

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