玉米InDel标记20重PCR检测体系的建立

Establishment of Multiplex PCR System with 20 Pairs of InDel Markers in Maize

为构建玉米多重PCR检测体系,提高分子标记检测效率,利用10份代表性玉米材料对238对InDel引物进行单重PCR评估,共得到192对扩增效率高、稳定性好的引物。根据软件评估结果、扩增质量、产物范围、染色体均匀分布原则从192对引物中优选出30对综合表现较好的引物形成扩增产物范围在80~200 bp和200~400 bp的两组核心引物组合,每套组合中有10对引物分布在不同染色体上。在核心引物组合的基础上综合考虑染色体分布、碱基片段范围、引物荧光颜色,逐一添加引物,最终形成两组玉米20重PCR体系,一组40重荧光标记毛细管电泳。

In order to improve the detection efficiency with molecular marker,the multiple PCR detection system was constructed.In this study,10major materials were used to evaluate the single pair PCR with238pairs of InDel primers.According to thesoftware quality evaluation results,product range,and the principle of chromosome uniform distribution,30pairs of primers wereselected from192primers with better performance to form two groups of core primer combinations with amplified products in therange of80?200bp and200?400bp,There were10pairs of primers distributing in different chromosomes for each primer combination.Based on core primer combination and comprehensive consideration on chromosome distribution,base fragment,andprimers fluorescent color,we established two groups of corn test20PCR system and a group of40fluorescent capillary electrophoresis.