摘要
为建立杏适宜的ISSR反应体系,以红玉杏为试材,探讨影响ISSR扩增的主要因素包括模板浓度、MgCl2浓度、dNTPs浓度、引物浓度及循环次数。通过筛选及优化,确定杏ISSR的适宜扩增条件。结果表明,在25μLPCR反应体系中,各组分的适宜浓度配比为:模板1μL(40ng)、dNTPS2.5μL(2.5mmol/L)、TaqDNA聚合酶0.5μL(2.5U)、引物1μL(20μmmol/L)、10×PCRBuffer2.5μL、MgCl22.5μL(2.5mmol/L)、ddH2O15μL,反应循环次数40次。同时利用建立的ISSR反应体系对3种普通杏品种进行扩增,证明该体系完全适合此标记对杏不同品种的遗传分析,从而为下一步杏资源遗传多样性的研究提供理论依据和参考价值。
The main factors affected ISSR reaction were tested with Hongyu apricot as test materials to optimize ISSR amplification system,including the concentration of template,MgCl 2,dNTPs and primer concentration as well as cycle numbers.The suitable ISSR amplification conditions of apricot were then conducted through the screening and optimization.The results showed that the optimal ISSR reaction was carried out in a volume of 25 μL containing 1 μL template DNA （40 ng）,2.5 μL dNTP S （2.5 mmol/L）,0.5 μL Taq DNA polymerase （2.5 U）,1 μL primer （20 μmmol/L）,2.5 μL 10×PCR Buffer,2.5 μL MgCl 2 （2.5 mmol/L）,15 μL ddH 2 O and 40 cycle numbers.The ISSR amplifications for 3 cultivars of common apricot were then investigated through the above reaction system.The system was completely suitable to analyze the genetic information for different apricot cultivars and thus supplied the further genetic diversity of apricot with theoretic bases and referenced value.
出处
《中国农学通报》
CSCD
北大核心
2010年第21期218-222,共5页
Chinese Agricultural Science Bulletin
基金
山东省自然科学基金"李梅杏种质资源的遗传多样性及利用研究"(Q2008D01)
关键词
杏
ISSR
优化
apricot （Prunus armeniaca L.）
ISSR
optimization
