河西走廊地区盐碱土壤微生物总DNA提取方法的比较

doi:10.13866/j.azr.2015.05.04

摘要/Abstract

摘要： 为了找出一种更适于河西走廊地区盐碱土壤微生物总DNA的提取方法,分别运用方法1(Omega试剂盒法)、方法2(MoBio试剂盒法)、方法3(CTAB-SDS法)和方法4(CaCl₂-SDS-酶解法),对河西走廊盐碱土壤微生物总DNA的提取方法进行比较。4种方法所提取总DNA片段分子量大约在23.1 kb,完整性较好,无明显降解现象。但4种方法所提取DNA的量差异很大,其中方法1和方法2提取的DNA目的条带不清晰,但纯度较高;方法3和方法4所提取的DNA产量高于方法1和方法2,但方法3提取的总DNA杂质较多,影响后续分析的准确性;而方法4所提取得DNA纯度基本与试剂盒法接近,DNA提取量可达到452.875 μg·g^-1,远远高于其他3种方法,能满足后续实验,是一种较理想的且适合于河西走廊盐碱土壤微生物多样性和种群结构的总DNA提取方法。

关键词: 盐碱土壤, 微生物, 总DNA提取方法, 河西走廊

Abstract: Due to soil microbial community structure and diversity and their change to a certain extent reflect the quality of the soil,the saline soil microbial diversity and population structure have certain application value for saline soil improvement. Saline soil is a special kind of soil type,may be a lot of salt resistance,alkali resistance and halophilic extremophiles groups,and has great development and utilization value,can build saline soil microbial macro gene pool,and screen out useful genetic resources used by mankind. To establish a method of total DNA extraction for investigation of the microbial population structure and diversity at saline-alkali soil in Hexi corridor,four kinds of methods that are M1(Omega kit method ),M2(MoBio kit method ),M3(modified high-salt method) and M4(Calcium chloride-SDS-Enzymatic) were used. The results showed that: 23.1 kb DNA fragments were obtained by the methods,and no DNA degradation. But the DNA extraction results were very different between the 4 methods,M1 and M2 shows no clear bands but high pure DNA; M3 and M4 obtain higher DNA amount than M1 and M2,however,the DNA obtained by M3 has more impurities which affects the accuracy of subsequently analysis. The DNA purity by M4 are more approximate to the kit method,the amount of DNA extraction reached 452.875 μg·g^-1,much higher than the other 3 kinds of methods,more suitable for the subsequent experiments. The results indicated that M4 is suitable for total DNA extraction to study microbial abundance and diversity from saline-alkali soil in Hexi Corridor.

Key words: saline-alkali soil, microorganism, total DNA extraction method, Hexi Corridor

李海云, 牛世全, 孔维宝, 王蓓, 达文燕. 河西走廊地区盐碱土壤微生物总DNA提取方法的比较[J]. 干旱区研究, 2015, 32(5): 856-860.

LI Hai-yun, NIU Shi-quan, KONG Wei-bao, WANG Bei, DA Wen-yan. Comparison of Microbial Total DNA Extraction Methods from Saline-alkali Soils in Hexi Corridor[J]. Arid Zone Research, 2015, 32(5): 856-860.

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