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作 者:赵宏[1] 王省芬[1] 张桂寅[1] 马峙英[1]
出 处:《河北农业大学学报》2002年第1期1-4,共4页Journal of Hebei Agricultural University
基 金:河北省自然科学基金 (30 116 7);河北农业大学重点科研基金 (9816计划 )资助项目
摘 要:对影响棉花AFLP技术体系的多种因素作了探讨 ,得到了一种适于棉花AFLP银染技术的优化体系。该体系中各优化因素为 :①模板DNA浓度为 15 0ng·μL-1;②酶切体系中 ,MseI和EcoRI各加入 3units ,使用NEBbuffer2 ,反应时间为 2h ;③连接最适反应时间为 10h ;④连接产物最适稀释倍数为 10倍 ;⑤预扩增产物最适稀释倍数为10倍。In this paper,some factors affecting the AFLP researching system of cotton were studied and an advanced technical system was established. The ascer tained factors in the system are 1) concentration of template DNA is 150 ng·μL -1 . 2) 3 units of MseI and EcoRI are added into digest system with NEB buffer 2 and 2 hours is enough to reaction. 3)the time of ligation should be 10 hours at least. 4) the products of ligation are diluted to 10 times with 1XTE for preamplification. 5) the products of preamplification are diluted to 10 times with 1XTE for selective amplification.
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