芒果SRAP-PCR反应体系的优化与退火温度的筛选

SRAP-PCR OptimizationandAnnealing Temperature Selection for Mango

以芒果为材料,通过单因子模板DNA含量、引物含量、2×Taq PCR Master Mix含量3种因素水平进行试验,建立了SRAP反应的优化体系。结果表明:最佳反应体系为20μL,包括24ng模板DNA,8pmL引物,2×Taq PCR Master Mix 10.0μL。该体系对芒果扩增效果好,电泳结果显示扩增条带清晰,多态性高,而且操作简单。从81对引物组合中筛选出扩增条带清晰、多态性丰富的29对引物组合。在此基础上对退火温度进行探索,发现扩增结果对退火温度变化不敏感。

Mango seedlings were used as the test material in this experimentation to optimize SRAP by the single factor tests of primer DNA ,primer ,and 2xTaq PCR Master Mix . The results indicated that a 20 uL reaction system appeared to be most desirable ,as the resultant electrophoresis provided clear amplification bands and was highly polymorphic while the procedure was simple .From 81 primer combinations ,29 were selected with abundant polymorphism .The optimized SRAP-PCR system and polymorphism primer combinations could be applied for molecular genetic studies on mango .Our investigation showed that annealing temperature was not a significant factor on the amplification .