固始鸡与安卡鸡F_2资源群Myf5基因PCR-SSCP分析条件的优化  

Optimization of PCR-SSCP Conditions for Analysis of Gushi-Anka Crossing F_2 Generation Chickens Myf5 Gene

作　　者：黄煌[1] 马新红[1] 康相涛[1,2] 韩瑞丽[1] 孙桂荣[1] 亢娟娟[1] 刘凯[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]河南省家禽种质资源创新工程研究中心,河南郑州450002

出　　处：《家畜生态学报》2010年第1期34-37,共4页Journal of Domestic Animal Ecology

基　　金：国家科技支撑计划子课题(2006BDA01A09);国家蛋鸡产业技术体系项目资助;公益性行业(农业)科研专项子课题(ny-hyzx07-039);河南省杰出人才创新基金(074200510009)

摘　　要：为进行固始鸡与安卡鸡F2资源群Myf5基因的多态性研究,建立适合于该资源群的Myf5基因PCR-SSCP分析方法,对影响单链构象多态性(SSCP)图谱分辨率的凝`胶浓度、甘油浓度、电泳电压、电泳温度、电泳时间、PCR产物与上样缓冲液的比例、电泳缓冲液浓度、上样量等影响因素进行了比较系统的优化试验研究。结果表明:凝胶浓度10%、甘油浓度50%、上样量10μL、PCR产物与变性缓冲液比例为10∶10、电泳缓冲液为1×TBE、室温下300 V预电泳10 min后120 V电压电泳16 h,染色后条带细而清晰,能够准确分辨各基因型,为试验SSCP分析的最佳条件组合。In order to establish proper conditions of polywerase chain reaction-single strand conformation polymorphism（PCR-SSCP） for studies in Gushi-Anka Crossing F2 Generation Chickens Myf5 gene,an optimization test was carried out on influential factors including gel concentration,glycerol concentration,voltage,temperature,time,proportion of PCR product to loading dye,ionic strength of runing buffer,loading amount etc.The results showed that the optimal experiment conditions were as follows： glycerol concentration of 50%,gel concentration of 10%,PCR product loading amount 10μL,10∶10 of PCR product to loading dye,running buffer of 1.0 ×TBE,and pre-voltage of 300 V 10 min,then 200 V electrophoresis for 16 hours at room temperature.With above optimized conditions,the best PCR-SSCP method for analysis of the polymorphism in Gushi-Anka Crossing F2 Generation Chickens Myf5 gene was established.

关 键 词：固始鸡与安卡鸡F2资源群 Myf5基因 PCR-SSCP 正交试验 分析条件优化 

分 类 号：S811.5[农业科学—畜牧学]