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尽管人们采用多种提取方法乃至使用商品试剂盒,但从森林土壤中获得纯品DNA仍然是比较困难的事情。本文经多次研究探索提出了一种经济有效的森林土壤粗品DNA进一步纯化方法。此方法由两步组成:1.利用提取缓冲液将提取的粗品DNA溶解,经氯仿/异戊醇去除杂质后,再用异内醇沉淀析出DNA;2.使用普通凝胶回收试剂盒回收柱进一步纯化DNA。结果表明:经第一步纯化后,82%-91%腐殖酸杂质被除去。经第二步纯化后,残留的少量腐殖酸杂质被去除干净。经上述连续的两步纯化后获得的DNA非常纯净,可直接用于对抑制物非常敏感的常规PCR反应。本研究报道的DNA进一步纯化方法有效、经济且省时。此外,采用其它各种提取方法获得的土壤粗品DNA均可使用本方法进一步纯化。图4表2参15。
Although people use many extraction methods and even commercial kits, obtaining pure DNA from forest soils is still more difficult. This paper, after many studies, explored and proposed a cost-effective method for the further purification of crude DNA from forest soils. This method consists of two steps: 1. The extracted crude DNA is dissolved in an extraction buffer, and the DNA is precipitated by iso-internal alcohol after chloroform / isoamyl alcohol is removed; 2. The column is recovered using an ordinary gel recovery kit Further purify the DNA. The results showed that after the first purification, 82% -91% of humic acid impurities were removed. After the second purification, a small amount of residual humic acid impurities are removed. The DNA obtained after the above two successive purification steps is very pure and can be used directly in conventional PCR reactions that are very sensitive to inhibitors. The DNA purification methods reported in this study are effective, economical and time-saving. In addition, the crude soil DNA obtained by various other extraction methods can be further purified by this method. Figure 4 Table 2 参 15.