利用荧光AFLP技术检测罂粟DNA多态性

DNA Polymorphism Detection of Papaver Somniferum L Using Fluorescent Amplified Fragment Length Polymorphism

目的利用荧光AFLP检测技术检测罂粟植物DNA多态性。方法用Axygen公司的AxyPrep DNA试剂盒提取了12株产于缅甸和中国云南省昆明市宜良县罂粟植株的DNA,用EcoRI和MseI对总DNA进行酶切,连接人工接头,预扩增和选择性扩增,其产物在CEQ8000遗传分析系统上检测。结果64对选择性扩增引物中8对引物能得到20条以上的扩增片断,具有高度多态性。结论荧光AFLP技术可用于罂粟DNA多态性的检测。

Objective To detect DNA polymorphism of Papaver somniferum L using fluorescent Amplified Fragment Length Polymorphism. Methods Genomic DNA was isolated using the AxyPrep DNA Kit, doubledigested by two restrictional endonucleases （Eco RI and Mse I） and ligated to oligonucleotide adapters. After Pre-amplification and selective amplification, the DNA fragments were separated by capillary electrophoresis using the CEQ8000 DNA Fragment Analyzer. Results More than 20 fragments of highly polymorphic products were obtained by 8 pairs of primer from 64 selective amplifying primer pairs. Conclusion The fluorescent AFLP technique can be used to detect the DNA polymorphism of Papaver somniferum.