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Xu Z.-G.,Chaohu Vovational and Technical College | Cai Y.-H.,Chaohu Vovational and Technical College | Yuan X.,Chaohu Vovational and Technical College | Liu X.-S.,Chaohu Vovational and Technical College | Jiang C.-J.,Chaohu Vovational and Technical College
Chinese Journal of Biologicals

Objective: To develop an improved method for extraction of genomic DNA from bacillus, which can be used directly for PCR. Methods Genomic DNAs were extracted from Bacillus cereus, Bacillus thuringiensis and Bacillus natto by cetyl trimethyl ammonium bromide (CTAB)-lysozyme-frozen-thaw lysis method and determined for Avalues at wavelengths of 230, 260 and 280 nm by utralviolet spectrophotometer, based on which the DNA concentrations of extracted samples were calculated. The extracted DNAs were used as templates for PCR. Results The A260/A280, values of DNAs extracted by the improved CTAB method were 1.8-2.0, while the A260/A230 values were more than 2.0, and the DNA concentrations were more than 110 μg/ml. PCR product showed clear bands each at length of 1 500 bp and a high concentration, while no other specific bands were observed, and all the bands were clear. Conclusion: CTAB-lysozyme-frozen-thaw lysis method was simple and effective for extraction of genomic DNA from bacillus and might be used for PCR, which was suitable for molecular biological test in clinic. Source

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