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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。
A variety of factors affecting cotton AFLP technology system were discussed, and an optimized system for cotton AFLP silver staining technology was obtained. The optimization factors in this system are as follows: ① template DNA concentration is 15 0 ng · μL-1; ② In enzyme digestion system, 3 units are added to MseI and EcoRI, and reaction time is 2 hours with NEBbuffer2; ③ the optimal reaction time is 10 hours; ④ the optimal dilution of the product was 10 times; ⑤ pre-amplification of the product of the optimum dilution of 10 times.