摘要
本文以苹果为试材，研究了影响苹果ISSR—PCR反应的相关因素，并筛选了苹果ISSR—PCR引物。结果表明，在25μL ISSR-PCR反应体系中，适宜的引物和模板浓度分别为0．8μmol／L和20ng。适宜的退火温度可减少非特异扩增，提高PCR产物的多态性和差异性，退火温度大部分比Tin值低2-4℃。从112个引物中筛选出17个多态性较好的引物，共扩增出113个位点，其中多态性位点74个，所占比例为65．5％。
Influencing factors which affected ISSR-PCR in apple were studied and primers selected in this study. The results showed that 25 μ 1 reaction volume including 0.8 μ mol/L primer and 20ng DNA was proper in ISSR-PCR of apple. The suitable annealing temperature could reduce non-specific amplification and improve polymorphism and differences in PCR product. Most of the annealing temperature was 2-4℃ lower than Tin. 17 primers were selected from 112 ISSR primers, and a total of 113 DNA bands were ampli-fied, 74 of which （65.5%） were polymorphic.
出处
《吉林农业科学》
CSCD
北大核心
2013年第6期68-70,89,共4页
Journal of Jilin Agricultural Sciences
基金
吉林省科技发展计划项目(20080239)
