摘要： 以山葡萄品种左山一的叶片和花芽为材料,对cDNA-AFLP体系的几个关键因素进行摸索,建立了适宜山葡萄的cDNA-AFLP反应体系.结果表明,提取山葡萄的花芽RNA适宜用改良的CTAB法,提取叶片RNA适宜用SDS-酸酚法.在2x Y/T Buffer的作用下,500 mg的cDNA利用MseⅠ与EcoRⅠ双酶切4 h即可酶切完全,将16℃连接过夜的产物稀释倍作为预扩增的模板,预扩增产物稀释20倍作为选扩模板能获得带型稳定可重复性好的结果.

关键词: 山葡萄, cDNA-AFLP

Abstract: The cDNA-AFLP systemfor Vitis amurensis Rupr was established after optimizing several key factors affecting cDNA-AFLP analysis.The result indicated that CTAB method could be used for extracting ideal RNA from leaf and inflorescence of Vitis amurensis Rupr,while SDS-acidic phenol method for extracting RNA from bud;500 ng cDNA was digested completely by EcoR I and Mse I for 6 hours.The reducible results was obtained when the ligation products were diluted to 5 times for preamplification template and preamplification productsthe were diluted to 20 times for selective amplification.

Key words: Vitis amurensis Rupr, cDNA - AFLP

中图分类号: 

• S663.1