摘要
本研究以19份薰衣草品种为材料,采用正交设计对ISSR-PCR反应中的5个影响因素：DNA浓度、Taq DNA聚合酶用量、Mg2＋浓度、引物浓度、d NTP浓度,进行5个水平的优化试验,并在48℃~59℃范围内摸索退火温度。研究结果建立了稳定的、可重复的薰衣草ISSR最佳反应体系和PCR扩增参数。在20μL的反应体系中,DNA模板浓度为2.50 ng/μL,Mg2＋浓度为2.00 mmol/L,d NTP浓度为0.20 mmol/L,引物浓度为0.40μmol/L,Taq DNA聚合酶为0.75 U,退火温度为52℃。该体系可为开展薰衣草种质资源的遗传多样性分析评价奠定基础。
The experimental designs with orthogonal test was used to optimize the ISSR-PCR reaction procedures of nineteen lavender cultivars in five factors including the concentration of DNA, Taq DNA polymerase, d NTP, primers and Mg2＋at five levels respectively. Then, the annealing temperature of the primers were researched in 48℃ ~59℃ as well. A stable and repeatable reaction system of ISSR-PCR for lavender was established, PCR reaction volume of 20 μL included 2.50 ng/μL, DNA template, 2.00 mmol/L Mg2＋, 0.20 mmol/L d NTPs, 0.40 μmol/L primer concentration and 0.75 U Taq DNA polymerase dosage and proper annealing temperature was 52℃. The establishment of the PCR reaction conditions could settle favorable basis for the further study on the genetic diversity of Lavandula Germplasm by using ISSR molecular marker techniques.
出处
《分子植物育种》
CAS
CSCD
北大核心
2015年第3期641-645,共5页
Molecular Plant Breeding
基金
新疆维吾尔自治区高校科研计划青年教师科研启动基金(XJEDU2012S13)
国家自然科学基金(31460383)
新疆农业大学校前期课题(XJAU201017)共同资助
作者简介
通讯作者，smm1980@yeah．net
