大薯ISSR-PCR反应体系的优化

Optimization of ISSR-PCR Reaction System for Dioscorea alata L.

以大薯为材料,通过试验优化了ISSR-PCR反应体系中的主要成分用量及浓度、退火温度对大薯ISSR扩增结果的影响。试验表明:在20μL的反应体系中,Mg2+的浓度为1.875 mmol/L,dNTPs浓度为0.500 mmol/L,引物的浓度为1.0μmol/L,TaqDNA聚合酶的用量为1U,模板DNA的用量为60 ng,在48℃的退火温度下45个循环,能得到清晰、多态性高的ISSR带谱。

The factors that affecting the ISSR（inter-simple sequence repeat）result of Dioscorea alata L.were studied.By adjusting DNA template,primer,Mg2＋,dNTPs,Taq DNA polymerase,annealing temperature and cycle of PCR reaction were optimized.The results showed that the optimized content consists of DNA was 60 ng,primer was 1.0 μmol/L,Mg2＋ was 1.875 mmol/L,dNTPs was 0.500 mmol/L,Taq DNA polymerase was 1U,annealing temperature was 48℃,45 cycles of PCR reaction,in 20 μL reaction system.