玉竹ISSR反应体系的优化  

Optimization for reaction system of Polygonatum odoratum ISSR

作　　者：陈玉秀[1] 周日宝[1] 潘清平[1] 姜傲芳[2] 吴佳[1] 童巧珍[1] 

机构地区：[1]湖南中医药大学药学院,湖南长沙410007 [2]中南林业科技大学经济林育种与栽培国家林业局重点实验室,湖南长沙410004

出　　处：《湖南中医药大学学报》2008年第2期37-39,共3页Journal of Hunan University of Chinese Medicine

基　　金：国家科技支撑项目(2006BAI06A11-06)

摘　　要：目的建立稳定性高、重复性好的适合玉竹ISSR-PCR扩增的反应体系。方法以玉竹根茎为材料,采用改良的CTAB法提取玉竹基因组DNA,并利用单因素多水平试验,分别对模板DNA、引物、Mg2+、Taq酶、dNTP的浓度和退火温度进行考察。结果玉竹的最佳反应体系为:在20μL的反应体系中,含1×buffer、15 ng模板DNA、0.4μmol/L引物、2 mmol/L Mg2+、1.0 U Taq酶、200μmol/L dNTP。此反应体系以52℃的退火温度扩增出来的玉竹电泳图谱清晰,重复性好。结论本实验反应体系适宜于玉竹ISSR-PCR的扩增,为玉竹下一步分子生物学的研究打下基础。Objective To establish a high stabilizing reaction system for Polygonatum odoratum ISSR-PCR. Methods The Polygonatum odoratum DNA was extraeted with improved CTAB technique and the template DNA concentration, primer concentration, Mg^2＋ concentration, dNTP concentration, Taq polymerase dosage and temperature were researched respectively. Result The best reaction system of Polygonatum odoratum was in the 20 μL system containing lxbuffer, 15 ng template DNA, 0.4 μmol/L primer, 2 mmol/L Mg^2＋, 1.0 U Taq polymerase, 200 μmol/L dNTP. Conclusion This reaction system is suitable to amplification of ISSR-PCR of Polygonatum odoratum and has given good foundation for further study of Polygonatum odoratum.

关 键 词：玉竹 简单重复序列间 反应体系优化 

分 类 号：R284.2[医药卫生—中药学]