摘要
以黑珍珠和砂蜜豆为材料,对AFLP分析过程中模板DNA的制备、酶切与连接、酶切连接产物的预扩增与选择性扩增、聚丙烯酰胺凝胶电泳等过程中易出现的问题及其解决方法作了对比研究,建立了一套适合于甜樱桃的AFLP分子标记体系。共筛选了64对引物,其中30对扩增效果理想,能够得到清晰、稳定的条带,平均每对引物扩增条带为60条。该体系的建立为研究甜樱桃遗传多样性和标记农艺性状的技术平台构建奠定了基础。
The variety Heizhenzhu and Shamidou were used as materials for establishing DNA-AFLP system. Sev- eral key factors affecting DNA-AFLP analysis were detected such as DNA extraction, enzymes restriction, adaptor-liga- tion,pre-amplification and selective amplification. The results indicated that SDS method could be used for extracting ideal DNA from leaf of sweet cherry;500 ng DNA was digested completely by EcoR I and Mse I for 4 h. The reducible results was obtained when the ligation products were diluted to 5 times for pre-amplification template and pre-amplifi- cation products were diluted to 20 times for selective amplification. This study established a foundation for studying the cherry genetic diversity and QTL for cherry agricultural character.
出处
《华北农学报》
CSCD
北大核心
2010年第2期125-128,共4页
Acta Agriculturae Boreali-Sinica
基金
国家自然科学基金资助项目(506090022)
