随机扩增多态DNA规范化反应体系的探讨  

Researdl on the Standard Condition of RAPD Analysis in Pigs

作　　者：邓昌彦[1] 蒋曹德[1] 

机构地区：[1]农业部猪遗传育种重点开放实验室,武汉430070

出　　处：《黄冈职业技术学院学报》2002年第4期52-55,共4页Journal of Huanggang Polytechnic

基　　金：国家"973"项目(G2000016105)

摘　　要：在研究猪分子标记遗传距离同杂种优势的关系时,对模板浓度和纯度、引物浓度、dNTP浓度、Mg2+浓度、不同商标的Tag酶等影响RAPD扩增的因素进行了系统的分析,在此基础上建立了适宜于本实验室研究的最佳RAPD技术体系:25(1反应体系中,含10×buffer2.5(I,MgCl21.75mmol/L,dNTPO25mmol/L,引物0.24(mol/L,Tag酶2U,模板50～100ng,1(g/(IBSA。反应程序为:94℃预变性5min,然后94℃变性1min,36℃退火1min,72℃延伸2min40个循环,最后在72℃延伸10min。In order to research the relationship between genetic parents distance and heterosis in pigs, the authors probe to some essential factors affecting RAPD pattern, including template concentration and purity, primer concentration, dNTP concentration, Mg+ + concentration, different brand Tag polymerase, etc. The optimal condition of amplification for RAPD assay in pigs as follows: in 25(1 reaction solution, containing 10 × buffer2.5(1,MgC121.75mmol/L,dNTP0.25mmol/L, random primer0.24(mol/L, DNA polymerase2U,BSAl (g/(1. The PCR profiles are 5min at 94℃ for 1 cycle followed by 1 rain at 94℃, 1min at 36℃ ,2min at 72℃ for 40 cycles and 10 min at 72℃ as final extension step.

关 键 词：猪 RAPD 影响因素 规范化 

分 类 号：S828.2[农业科学—畜牧学]