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 :2 5 μl反应体系中 ,含 10×buffer 2 .5 μl,MgCl2 1.75mmol/L ,dNTP 0 .2 5mmol/L ,引物 0 .2 4μmol/L ,Tag酶 2U ,模板 5 0～ 10 0 μg ,1μg/μlBSA。反应程序为 :94℃预变性 5min ,然后 94℃变性 1min ,36℃退火1min ,72℃延伸 2min4 0个循环 ,最后在 72℃延伸 10min。
In order to research the relationship between genetic parents distant 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 25μl reaction solution, containing 10×buffer 2.5μl,MgCl 2 1.75mmol/L,dNTP 0.25mmol/L,random primer 0.24μmol/L, DNA polymerase2U,BSA1μg/μl. The PCR profiles are min at 94℃ for 1 cycle followed by 1 min at 94℃,1min at 36℃,2min at 72℃ for 40 cycles and 10 min at 72℃ as final extension step.
出处
《生物技术通报》
CAS
CSCD
2002年第3期40-43,共4页
Biotechnology Bulletin
