鞣制皮张DNA的提取

DNA Isolation from Tanned Animal Skins

采用5种方法从鞣制的野猫(Felissilvestris)皮和漠猫(F.bieti)皮中提取DNA,用Cytb基因的通用引物进行PCR扩增,并对扩增结果进行测序,以检验提取效果。结果显示,鞣制皮张中的DNA含量少,高度降解,且含有大量的酶抑制物。用酚-氯仿法可以得到DNA,但需要采用DNAIQTM系统(Promega)的磁珠纯化策略进一步彻底纯化才能达到最佳扩增效果。此外,PCR引物设计时应尽量缩短扩增片段长度以便提高扩增成功率。

Tanned skins are valuable materials for molecular genetic study of wildlife due to their abundance in species and number, multiple sources and long preserving capacity. However, tanned skins have not been widely used for that purpose because of a lot of technical restrictions in DNA isolation. Five methods were tested to isolate DNA from tanned skins of wild cat （Felis silvestris） and Chinese desert cat （F. bieti）. In order to test the effectiveness of these methods, three universal primer pairs of Cytb gene were used for PCR amplification and PCR products were directly sequenced. Results indicate that tanned skins still withhold a tinny amount of but highly degraded DNA together with considerable enzyme inhibitors. DNA can be effectively isolated by using conventional phenol-chloroform method, but requires thorough purification using the means of magnetic bead strategy provided by DNA IQTM system （Promega）. In addition, the length of amplicons should be reduced to fit the fragment size of isolated DNA template when primers are designed.