摘要
本研究采用L16(45)正交试验设计,对油用牡丹ISSR反应体系中的5个主要因素进行了优化,确立了ISSR-PCR的最佳反应体系:反应体系总体积20μL,含0.50 U Taq聚合酶,20 ng模板DNA,3.00 mmol/L Mg^(2+),0.40 mmol/L d NTPs,1.00μmol/L引物及10×PCR Buffer。基于ISSR-PCR最佳反应体系,我们共筛选出了12条多态性丰富、条带清晰稳定的引物,为油用牡丹遗传连锁图谱的构建及分子育种奠定基础。
The orthogonal experiment design L16（45） was used to optimize the five major factors in oil tree peony ISSR reaction system, and the optimized ISSR-PCR reaction system for oil tree peony was also established. The optimal ISSR-PCR reaction system was as follows： total volume 20 μL, containing 0.50 U Taq DNA polymerase,20 ng template DNA, 3.00 mmol/L Mg2＋, 0.40 mmol/L d NTPs, 1.00 μmol/L primer, and 10 ×PCR Buffer. 12 primers with rich polymorphisms and clear bands were screened by the optimal ISSR-PCR reaction system, which will provide the basis for the genetic linkage map and molecular breeding for oil tree peony.
出处
《分子植物育种》
CAS
CSCD
北大核心
2017年第8期3129-3135,共7页
Molecular Plant Breeding
基金
上海市绿化和市容管理局重点科技攻关项目(F122431
G162403)
上海市科学技术委员会自然基金(11ZR-1436100)共同资助
