摘要
通过正交试验建立芦笋ISSR优化反应体系,并对其反应程序进行优化。实验结果表明,优化的扩增体系:25μL的反应体系中含150 ng的模板DNA、0.3μmol/L引物、15μL 2×Mastermix;优化的反应程序:94℃预变性5 min;94℃变性45 s,设定温度退火60 s,72℃延伸45 s,循环30次;然后72℃延伸10 min,4℃保存;筛选到了21条引物并确定其最优退火温度。实验结果为芦笋资源遗传多样性评价的研究奠定基础。
The optimum ISSR reaction system in A.officinalis was established by orthogonal test,then its reaction process was optimized.The result showed that the optimized amplification system was 150 ng temple DNA in the total volume of 25 μL ISSR system,0.3 μmol/L primer,and 15 μL 2× Mastermix.The optimized PCR procedure was： pre-denaturation at 94℃ for 5 min,denaturation at 94℃ for 45 s,setting temperature annealing for 60 s,extension at 72℃ for 45 s,and repeated for 30 cycles; then extension at 72℃ for 10 min,and remaining at 4℃.Based on this optimized system,21 primers were screened and the optimal annealing temperature was set.The results could lay the foundation for the genetic diversity evaluation of A.officinalis.
出处
《分子植物育种》
CAS
CSCD
北大核心
2018年第1期170-174,共5页
Molecular Plant Breeding
基金
海南省应用技术研发与示范推广专项(ZDXM2015047)
公益性行业(农业)科研专项经费项目(201003074)共同资助
关键词
芦笋
ISSR
体系优化
Asparagus officinalis
ISSR
System optimization
