TP-M13自动荧光检测法在高粱SSR基因型鉴定中的应用

研究利用TP-M13自动荧光检测法对48份高粱材料进行了简单序列重复(SSR)标记的基因型鉴定.在这个方法中,需要合成一个普适性的用荧光(如FAM)标记的M13引物,并把M13的正向引物和一个SSR反向引物相连(称为TP-M13引物),利用3条引物序列进行PCR扩增,其PCR产物在DNA测序仪(如ABI3700仪)上进行自动荧光检测.结果表明,这种方法和其他的传统方法相比,具有经济、灵敏、高效的优点.建议在利用数量很多的SSR标记对数量有限的基因组较小的材料进行基因型鉴定时,使用TP-M13自动荧光检测系统.

Application of the TP-M13 Automated Fluorescent-lablled System of SSR Genotyping in Sorghum

Forty-eight sorghum accessions were genotyped with 104 SSR markers using the TP-M13 automated fluorescent-labelled system. In the method, one reverse SSR primer was tailed with a M13 forward primer, which is called TP-M13, the second SSR primer was normal, and the third primer was the M13 forward primer labeled by FAM. The PCR fragments amplified by these three primers were detected using an automated fluorescent-labelled system (DNA sequencer such as ABI3700). The result indicated that the method had the advantages of high-throughput, sensitiveness, cost-effectiveness and high accuracy over some of other conventional methods. Therefore, the TP-M13 automated fluorescent detection system is suggested to be suitable for genotyping limited number of accessions by the use of a number of SSR markers.