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目的 :筛选出最佳模板提取方法用于随机扩增多态性DNA(RAPD)。方法 :对 4种方法 (挑丝法、沉淀法、加热裂解法和裂解剂裂解法 )提取的模板DNA进行琼脂糖凝胶电泳、紫外线扫描及随机扩增多态性DNA ,比较其片段大小、纯度及RAPD指纹图差异。结果 :挑丝法和沉淀法均可提取出大于 2 3kb且纯度较高的模板DNA ,二者指纹图完全一致且均有 2～ 4kb的较大片段。加热裂解法和裂解剂裂解法提取的模板有弥散、碎裂的DNA并纯度较差 ,指纹图上仅有 6 0 0～ 2 0 0 0bp的较小片段。结论 :用挑丝法或沉淀法提取的模板DNA最适于进行RAPD。
Objective: To screen out the best template extraction method for random amplified polymorphic DNA (RAPD). Methods: The DNA extracted from the four methods (pick and draw method, precipitation method, pyrolysis method and lysis method) was subjected to agarose gel electrophoresis, ultraviolet ray scanning and random amplified polymorphic DNA. The fragment size, Purity and RAPD fingerprint differences. Results: Both the silkworm method and the precipitation method were able to extract template DNA with a purity of more than 23 kb and the purity was very high. The two fingerprints were exactly the same and all had large fragments of 2 ~ 4kb. The templates extracted by the pyrolysis method and the lytic reagent cleavage method have diffuse and fragmented DNA with poor purity and only have smaller fragments of 600 to 200 bp on the fingerprint map. CONCLUSION: The template DNA extracted by pick-and-place method or precipitation method is the most suitable for RAPD.