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高效电转化率条件的探索

Survey the Highest of Transformations-efficiency of the Competent Cell

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【作者】 张建琼; 张雪萍; 谢维; 单祥年; 林陵;

【Author】 ZHANG Jian qiong 1,ZHANG Xue ping 1,XIE Wei 2,SHANG Xiang nian 2,LIN Ling 1 (1 Department of Microbiology and Immunology,Nanjing Railway Medical College,Nanjing,210009,PRC) (2 Department of Biology,Nanjing Railway Medical College,Nanjing,21000

【机构】 南京铁道医学院微生物免疫学教研室!南京; 210009; 南京铁道医学院生物学教研室!南京;

【摘要】 对影响 DNA电转化效率的主要因素进行探索 .结果表明 :在相同的电场强度、电阻和电容条件下 ,电脉冲时间的长短对电转化效率有较大影响 ,电脉冲时间太长或太短均使转化效率降低 ;用对数生长早中期的细菌制备感受态细胞 ,经液氮速冻后储存于 - 70℃ ,能获得高转化效率 .

【Abstract】 Transformation-efficiency of the competent cell is one of the important factors that affects combinatorial library capacity.Some factors about transformation efficiency were compared and analyzed and the condition about the highest transforming efficiency was obtained.The present experiments demonstrate that the optical density at 600 nm of the E.coli XL 1 Blue is not more than 0.5,and electrocompetent XL 1 Blue placed in liquid nitrogen that they freeze instantaneously,and set up the Gene pulser to 2.5 kV,using a 25 uF capacitor and the pulse controller to 200 Ohm.Pulse once (press the button for two seconds after the signal tone) will obtain highly transforming efficiency.

【基金】 江苏省应用基础基金!资助 (BJ970 70 )
  • 【文献出处】 南京师大学报(自然科学版) ,JOURNAL OF NANJING NORMAL UNIVERSITY(NATURAL SCIENCE EDITION) , 编辑部邮箱 ,2000年02期
  • 【分类号】R392.11
  • 【被引频次】39
  • 【下载频次】878
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