湖北民族学院学报（自然科学版）2018，Vol.36Issue(2)：121-124,155,5.DOI:10.13501/j.cnki.42-1569/n.2018.06.001
华山松ISSR反应体系的优化
Optimization of ISSR Reaction System for Pinus armandii
摘要
Abstract
To obtain amplification result with clear,stable and rich polymorphism fragment of Pinus ar-mandii,the affection ISSR-PCR by main reaction system elements,such as primer,Mg2+and annealing temperature,was presented.The reaction system and amplified procedure suitable for P.armandii were es-tablished,the main elements include,15 μL amplification reaction mixture containing 1.5 μL 10×reaction buffer,22.5 ng template DNA,2.2 mmol/L Mg2+,0.23 mmol/L dNTPs,0.93 μmol/L primers,0.027 U/μL TaqDNA polymerase.The reaction program was devised for 4 min of predenaturalization at 94℃,33 cycles of 30 s for denaturalization at 94℃,30 s of anneal at Tm-4.8℃,108 s of extension at 72℃,and 10 min of extension at 72℃ after the final cycle.The 12 best primers were selected for ISSR-PCR system of P.ar-mandii from 44 primers,whose results of ISSR amplification were clear and stable.The research on the op-timization of ISSR reaction system for P.armandii will lay a foundation for studying interspecific genetic variation and genetic map construction of P.armandii using ISSR molecular marker technique in the future.关键词
华山松/ISSR/优化/PCRKey words
Pinus armandii/ISSR/optimization/PCR分类
农业科技引用本文复制引用
李翠新,何德..华山松ISSR反应体系的优化[J].湖北民族学院学报（自然科学版）,2018,36(2):121-124,155,5.基金项目
云南省应用基础研究计划项目(2008ZC092M) （2008ZC092M）
云南省教育厅科研基金研究生项目(2014J093) （2014J093）
云南省优势特色重点学科生物学一级学科建设项目(50097505). （50097505）
