利用几根猪毛进行PCR扩增以检测氟烷基因的基因型

Diagnosis of Genotypes for Halthane at DNA Level by a simple and Rapid Preparation of Hair for PCR Amplification

采集３０头日本某原种猪场杜洛克猪４根毛样，以简便的方法快速抽提ＤＮＡ，并进行ＰＣＲ扩增，得到特异的６５９ｂｐ的ＰＣＲ扩增产物，其中含有Ｃ１８４３或Ｔ１８４３的位点，经过ＨｈａⅠ和ＢｓｉｋｈａⅠ（相当于ＨｇｉＡⅠ）内切酶消化，得到酶切图谱，根据带型判断其发生变异与否。结果表明：①用猪毛进行ＰＣＲ扩增，方法简便，一次可大批量地处理样品，该方法是行之有效的。②用２～４根猪毛即可进行ＤＮＡ诊断，不需纯化ＤＮＡ。③在被检测的３０头杜洛克猪中，发现３头杂合子（Ｎ／ｎ），其余２７头为正常阴性纯合子（Ｎ／Ｎ）。

simple preparation using the root of plucked hairs for PCR amplification was established for diagnosis of genotypes for halothane in pigs.②As few as four hairs were required for the diagnosis.③Three out of 30 Duroc pigs were diagnosed as heterozygous carriers(Nn),and the rest pigs as halothane resistant(NN),using the above method.