藜ISSR-PCR反应体系的优化  

Optimization of ISSR-PCR system for chenopodium album L

作　　者：张恒庆[1] 朱立南[1] 杜洋[1] 贾俊玲[1] 谭琨[1] 

机构地区：[1]辽宁师范大学生命科学学院,辽宁大连116029

出　　处：《沈阳师范大学学报（自然科学版）》2011年第2期264-266,共3页Journal of Shenyang Normal University:Natural Science Edition

基　　金：辽宁省教育厅高校科研基金项目(2008353)

摘　　要：采用TaKaRa的PCR反应系统,以从藜(Chenopodium album L)幼叶中提取纯化后的总DNA为模板,进行UBC 859号引物的ISSR-PCR扩增实验,分别测试了镁离子、模板DNA和Taq DNA聚合酶含量对反应结果的影响,通过各因子的浓度梯度组合实验,确定了在25 mL体积的PCR反应中:酶配套缓冲液2.5μL,dNTPs0.2μmmol/L,引物0.2 mmol/L,Mg2+1.5 mmol/L,模板含量为40 ng,Taq DNA聚合酶为1.5 U是藜最佳的ISSR-PCR反应体系。In this study,the factors which affect the ISSR-PCR reaction were detected by TaKaRa PCR reaction system of Chenopodium album L.The purified template DNA was extracted from fresh young leaves,the effect of content of Mg2＋,templates DNA and Taq DNA polymerase were tested.According to the results of amplifications in gradient it was determined that the optimization ISSR-PCR reaction system in rolumes of 25 mL contained 2.5 μL to XPCR buffer,0.2 mmol/L mixture of dNTPs,0.2 mmol/L primer,1.% mmol/L Mg2＋,40 ng of DNA and 1.5 unit Taq DNA Polymerase.

关 键 词：藜 ISSR-PCR 反应体系 优化 

分 类 号：Q341[生物学—遗传学]