4种古DNA抽提方法效果比较

Comparisons of effects of four methods for ancient DNA extraction

目的：寻找到一种简便、快速、可靠、有效的古DNA抽提方法。方法：选择基于硅粒法和硅离心柱法的四种抽提方法（方法A：Modified Silica ParticlesMethod；方法B：Ceneelean Method；方法C：QIAamp Method；方法D：Modified QIAquick method））分别抽提5个距今约4000年的古绵羊线粒体DNA（mtDNA），并进行PCR扩增，通过琼脂糖凝胶电泳观察扩增成功与否，根据扩增成功率判断抽提方法的优劣。结果：4种方法的扩增成功率从20％～100％，基于硅离心柱法的抽提效果（60％，100％）明显的优于基于硅粒法（20％，40％），其中Modified Silica Particles Method的效果最差（20％），Modified QIAquick Method的效果最好（100％）。结论：Modified QIAquick Method方法使用Centrieon超滤管浓缩DNA抽提裂解液，结合硅离心柱进行DNA纯化，能有效去除PCR抑制物，同时得到较好的古DNA模板。

Objective To find a simple, rapid, reliable and effective method for ancient DNA extraction. Methods Four methods based on silica particles and silica-based spin columns （A： Modified Silica Particles Method; B： Geneelean Method; C： QIAamp Mothod; D： Modified QIAquick Method） were respectively used to extract mitochondrial DNA （mtDNA） from 5 ancient sheep dated to 4 000 years ago. PCR amplification of mtDNA was performed, and its results were evaluated by agarose gel electrophoresis. The effects of 4 kinds of methods mentioned above were determined by examining amplification success rates of DNA from 5 ancient sheep. Results The amplification success rates of 4 methods were different from 20% to 100~~, and the effects of extraction based on silicPbased spin columns （60%, 100%） were distinctly better than those based on silica particles （20~, 40%）. The effect of extraction based on modified silica particles was the worst, and based on modified QIAquick was the best among 4 methods. Conclusion Modified QIAquick Method is a highly effective method, with a 100% success rate, in which Centricone ultrafiltration devices are used to concentrate teeth digest solution and purify DNA through silica-based spin columns. It can effectively remove the inhibitor as well as obtain preferable DNA template.