革兰氏阳性与阴性细菌基因组DNA提取方法的优化

Optimization of Methods for Genomic DNA Extraction from Bacteria

在分析现有提取微生物基因组DNA的原理和方法基础上,对SDS-NaCl法进行了改良,以商品化微生物基因组提取试剂盒提取的基因组结果为对照,利用改良的SDS-NaCl法,分别提取大肠杆菌(Escherichia coli)、枯草芽孢杆菌(Bacillus subtilis)和金黄色葡萄球菌(Staphylococcus aureus)的基因组DNA。结果表明,改良的SDS-NaCl法提取的大肠杆菌、枯草芽孢杆菌和金黄色葡萄球菌基因组DNA的OD_(260)/OD_(280)分别为1.88、1.89、1.81,浓度分别为61.67、64.32、53.22μg/mL;而商品化试剂盒提取大肠杆菌、枯草芽孢杆菌和金黄色葡萄球菌基因组DNA OD_(260)/OD_(280)分别为1.85、1.87、1.83,浓度分别为64.07、58.43、52.34μg/mL。结果证明改良SDS-NaCl法提取的革兰氏阳性和阴性细菌的基因组DNA可用于后续试验如全基因组测序和PCR等,同时可快速获得大量的高质量微生物基因组。

Based on the analysis of the principles and methods of genomic DNA extraction,the SDS-NaCl method was modified.The results of genomic DNA extraction by commercial genomic extraction kit were compared with those by SDS-NaCl method modified by our laboratory.The results showed that the OD_(260)/OD_(280) of the genomic DNA of Escherichia coli,Bacillus subtilis and Staphylococcus aureus were 1.88,1.89 and 1.81,and the concentration were 61.67,64.32 and 53.22μg/mL respectively.The OD_(260)/OD_(280) by commercial genomic extraction kit were 1.85,1.87 and 1.83,and the concentration were 64.07,58.43 and 52.34μg/mL respectively.The results showed that the genomic DNA extracted by the modified nacl method could be used in subsequent experiments such as whole genome sequencing and PCR,and a large number of high quality microbial genomes could be obtained rapidly.