目的:比较不同方法提取苛求芽孢杆菌基因组DNA的差异。方法:用经典CTAB提取法、改进CTAB法(溶菌酶处理结合CTAB提取法)、UniQ柱吸附提取法制备苛求芽孢杆菌基因组DNA,比较产物完整性和用于PCR扩增的有效性。结果:三种方法制备基因组DNA纯度接近,但改进CTAB法产率最高,UniQ法产率最低。经典CTAB法和UniQ法提取基因组DNA易降解。三种方法所得基因组DNA用于PCR扩增效率接近。结论:溶菌酶裂解结合CTAB提取更适合制备苛求芽孢杆菌基因组DNA。
Objeetives:To compare methods for the preparation of genomic DNA from Bacillus fastidious. Methods: Genonic DNA was extracted by classical CrAB method, modified CTAB method with the use of lysozyme and UniQ ion- exchange method, the quality and quantity of genomic DNA were compared. Results: The modified CTAB method showed the highest yield, while the UniQ ion- exchange method showed the lowest yield of genomic DNA. These three methods gave DNA preparation of comparable homogenecity and efficiiency as the template for PCR amplification. Conclusion: The modified CTAB method was better for preparing genomic DNA from Bacillus fastidious.