绵羊MHC区段BAC克隆探针的制备  

Preparation of Probes Using the MHC Section BAC Cloning of Sheep

作　　者：邹毅辉[1] 李鑫[1] 李峰[1] 薛峰[1] 陈芳[1] 高剑峰[1] 

机构地区：[1]石河子大学生命科学学院,石河子832003

出　　处：《石河子大学学报（自然科学版）》2010年第6期700-702,共3页Journal of Shihezi University(Natural Science)

基　　金：国家科技部国际合作重大项目(2006DFB33750)

摘　　要：为了摸索适用于高通量核酸杂交筛选的探针的制备方法,采用GeneQuest软件分析并选取限制性内切酶BseDⅠ酶切MHC区段BAC克隆374N21,对酶切产物32P末端标记,并采用Omega E.Z.N.A DNA Probe Purifi-cation Kit纯化后放射自显影检测的方法。结果显示:酶切结果与软件分析一致,纯化后探针分布为0.5～2.0kb,纯化回收效率为60%。探针的分布合理,质量较高,可满足高通量杂交筛选的需要。In Order to find a way to manufacture probes that could meet the demand of high throughput hybrid screening,restriction enzymes of BseDⅠdigested the 374N21 clone of BAC MHC was analysed by using the Gene Quest software.The production of the digestion labeled with 32P,Autoradiography detection after being purified by Omega EZNA DNA Probe Purification Kit.The results showed that digestion results were consistent with the software analysis.Purified probes distribution in 0.5～2.0 kb.The efficiency of the purification recovery was 60%.The higher quality of probes can satisfy the needs of hybrid screening.

关 键 词：32P标记 绵羊 MHC 放射性自显影 DNA探针 

分 类 号：S826[农业科学—畜牧学] Q78[农业科学—畜牧兽医]