摘要
以辣椒花蕾和嫩叶为试验材料,对影响辣椒cDNA-AFLP体系的关键因素进行了探讨,建立了适于辣椒基因差异表达的cDNA-AFLP体系。结果表明,Trizol法适用于辣椒花蕾和嫩叶总RNA提取;SMART法合成双链cDNA效率较高;利用TaqI/AseI分步酶切cDNA各3h可酶切完全;连接产物最佳稀释倍数为15倍;预扩产物稀释15倍经选择性扩增,PAGE电泳可以获得带型丰富且重复性好的结果。辣椒cDNA-AFLP反应体系的建立为辣椒基因的差异表达分析奠定了基础。
The cDNA amplified fragment length polymorphism （cDNA-AFLP） analysis system of Capsicum annuum was established using buds and leaves as materials. The results indicated that Trizol method could be used for extracting ideal RNA from buds and leaves of pepper while double-stranded cDNA could be synthesized from total RNA by SMART method. Double-stranded cDNA was digested completely after 3h digestion by Taq I and Ase I step by step. The reducible result was obtained when the ligation products diluted to 15 times as templates for pre-amplification and 15 times-diluted pre-amplification products for selective amplification. The cDNA-AFLP analysis system was established successfully, which provided platform for further molecular study and gene cloning of Capsicum annuum.
出处
《中国农学通报》
CSCD
北大核心
2010年第12期26-29,共4页
Chinese Agricultural Science Bulletin
基金
国家自然科学基金(30370978)
仲恺农业工程学院引进优秀人才科研启动基金项目(G2360292)
