摘要
从27条简单重复序列区间(ISSR)引物中筛选出10条引物对19株酵母菌进行PCR扩增;采用正交试验设计方法,对ISSR的影响因素进行了研究,对Taq酶,Mg^2+,dNTP和引物进行了4因素3水平优化试验,再通过对退火温度和模板DNA浓度的筛选,得出ISSR—PCR扩增的最佳反应体系.10条引物共扩增出108条带,其中90.7%呈多态性,相似系数为0.48~1.00.
Ten primers selected from 27 ISSR primers were used for ISSR amplification of 19 tested yeast strains. The orthogonal design was used to optimize ISSR amplification system in four factors (Taq polymerase, Mg^2+ , dNTP, primer) at three levels respectively, ISSR amplification system was optimized in anneal temperature and template DNA concentration. A total of 108 bands were produced with 10 primers, among which 90.7 % were polymorphic, similarity coefficient ranged from 0.48 to 1.00.
出处
《浙江师范大学学报(自然科学版)》
CAS
2010年第1期89-94,共6页
Journal of Zhejiang Normal University:Natural Sciences
基金
浙江省自然科学基金资助项目(Y306172
Y3090343)
浙江省重大科技专项(2007C12036)
关键词
简单重复序列区间
酵母菌
正交设计
聚类分析
inter-simple sequence repeat (ISSR)
yeast
orthogonal design
cluster analysis