【摘要】 目的 :探讨大规模DNA测序技术平台中影响电转化效率的几个因素。方法 :电转化法测定经稀释或乙醇沉淀处理后连接反应混合物的转化效率。结果 :电转化电压在 2 .1～ 2 .5kV之间可获得高转化效率 ;乙醇沉淀法虽能降低盐浓度 ,但同时使微量质粒DNA丢失 ,不能提高电转化效率 ;稀释法能提高转化效率 5倍左右 ;连接反应物转化时 ,感受态细胞悬浮于无菌去离子水中的转化效率比悬浮于 10 %甘油无菌去离子水中的高。结论 :降低连接反应混合物的盐浓度能有效提高电转化效率 ,在大规模DNA测序技术平台中 ,急需研究开发一种纯化回收微量质粒DNA的简单而有效的方法更多还原

【Abstract】 Objective:To investigate some factors affecting electro transformation efficiency in large scale DNA sequencing platform.Methods:Electro transformation efficiency of the ligation mix was detected when the ligation mix was treated with the methods of water dilution or ethanol precipitation.Results:A high electro transformation efficiency was obtained when the voltage was set between 2.1 to 2.5 KV;ethanol precipitation would reduce the salt concentration but it could not increase the electro transformation efficiency because of the loss of the plasmid during the ethanol precipitation;water dilution increased the electro transformation efficiency by about five times;the electro transformation efficiency was higher when the competent cell suspended in deionized water than that in 10% glycerol.Conclusion:Lower salt concentration in the sample got a higher electro transformation efficiency and a simple and efficient method for purification of the ligation mix should be set up for the large scale DNA sequencing.更多还原