摘要
对蚕豆DNA提取质量和浓度、DNA双酶切与连接、酶切连接产物的预扩增和选择性扩增等AFLP技术体系中的关键技术进行了优化处理，构建了蚕豆AFLP银染技术体系。酶切与连接可在12，5μl体系中一步完成，酶切连接温度为37℃，反应时间12～14h；预扩增体系为20μl，选择性扩增体系为10μl。采用该技术体系应用8对引物构建的蚕豆种质资源AFLP指纹图谱，扩增条带多、多态性强且质量好，可满足遗传多样性分析要求。
Factors influencing faba bean AFLP analysis were studied, including the purification and concentration of genomic DNA, restriction and ligation of genomic DNA, the pre-amplification reaction and amplification reaction systems An optimized AFLP system of silver staining was established for genetic diversity analysis of faba bean germplasm. Accomplishment of restriction and ligation was in the same system at 37℃ , for 12h to 14h. The pre-amplification reaction system was 20μl and the selective amplification system was 10μl. It was shown that fingerprinting of faba bean constructed using 8 primer combinations, such as E-ACG/M-CTT, revealed sufficient amplified bands with high resolution and polymorphism.
出处
《植物遗传资源学报》
CAS
CSCD
2007年第2期153-158,共6页
Journal of Plant Genetic Resources
基金
中澳政府间大型双边合作项目(ACIARCS1/2000/035)
科技部植物种质资源共享平台建设(2003DEA3N024-07)
