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用几种方法抽提、纯化厌氧菌染色体DNA,并进行酶切分析,探讨有关反应条件及影响因素:(1)不同种属厌氧菌对溶菌酶裂解的敏感性不同,控制适当的反应体积及细菌裂解程度可提高溶菌酶法抽提的效率;对溶菌酶不敏感的厌氧菌可用碱裂解法抽提。(2)使用适当的氯化铯浓度,氯化铯密度梯度超离心法可较好地分离纯化DNA。(3)不同来源及结构的DNA,酶切条件不同;适当的亚精胺浓度可促进酶切反应。
Several methods were used to extract and purify anaerobic bacterial chromosomal DNA, and the enzyme digestion analysis was carried out to investigate the reaction conditions and influencing factors. (1) Different species of anaerobic bacteria have different sensitivity to lysozyme cleavage and control the appropriate reaction Volume and degree of bacterial lysis can increase the efficiency of lysozyme extraction; anaerobic bacteria that are not sensitive to lysozyme can be extracted by alkaline lysis. (2) The cesium chloride density gradient ultracentrifugation method can be used to separate and purify the DNA with a suitable concentration of cesium chloride. (3) Different sources and structures of DNA, different digestion conditions; the appropriate concentration of spermidine can promote the digestion reaction.