Gibson, D. G., Young, L., Chuang, R. Y., Venter, J. C., Hutchison, C. A. 3rd, & Smith, H. O. (2009). Enzymatic assembly of DNA molecules up to several hundred kilobases. Nature Methods, 6(5), 343-345.Gibson, D. G., Glass, J. I., Lartigue, C., Noskov, V. N., ...
从上图Nature Methods文章的摘要可看到,Gibson使用5‘核酸外切酶,DNA聚合酶和DNA连接酶完成了多DNA片段的一步连接,而这就是我们现在常用的Gibson assembly。 3. 实验过程示意 一般实验室都直接购买配好的Gibson assembly mixture,但也可自行购买T5 核酸外切酶、DNA聚合酶以及DNA连接酶配置。Gibson操作简单,具体过程和步...
An overview of Gibson Assembly, a technique to create exceptionally large molecular clones from PCR products
Gibson, D. G., Young, L., Chuang, R. Y., Venter, J. C., Hutchison, C. A. 3rd, & Smith, H. O. (2009). Enzymatic assembly of DNA molecules up to several hundred kilobases. Nature Methods, 6(5), 343-345. Gibson, D. G., Glass, J. I., Lartigue, C., Noskov, V. N.,...
Gibson, D.G. et al. (2010). Nature Methods. 901-903. Barnes, W.M. (1994). Proc. Natl. Acad. Sci.. 91, 2216-220. 操作说明、说明书 & 用法 操作说明 Gibson Assembly® Protocol (E5510) Gibson Assembly® Chemical Transformation Protocol (E5510) ...
通过核酸外切酶降解,切出粘性末端,两段DNA片段同源的粘性末端互补配对,再通过DNA聚合酶聚合、连接酶连接,便可将任意两段合适大小的DNA片段拼接起来(DanielGGibson,LeiYoung,Ray-YuanChuang,JCraigVenter,ClydeAHutchisonIII,HamiltonOSmith.EnzymaticassemblyofDNAmoleculesuptoseveralhundredkilobases.NatureMethods,2009,6(5)...
Reaction Summary- Gibson Assembly Reaction summary: Gibson, D. G., Young, L., Chuang, R. Y., Venter, J. C., Hutchison, C. A., 3rd, and Smith, H. O. (2009)Nature methods6, 343-345 Materials 1. 5X isothermal (ISO) reaction buffer (25% PEG-8000, 500 mM Tris-HCl pH 7.5, ...
两段DNA片段同源的粘性末端互补配对,再通过DNA聚合酶聚合、连接酶连接,便可将任意两段合适大小的 DNA 片段拼接起来(Daniel G Gibson, Lei Young, Ray-Yuan Chuang, JCraig Venter, Clyde A Hutchison III, Hamilton 0 Smith.Enzymatic assembly of DNAmolecules up to several hundred kilobases.Nature Methods, ...
两段DNA片段同源的粘性末端互补配对,再通过DNA聚合酶聚合、连接酶连接,便可将任意两段合适大小的 DNA 片段拼接起来(Daniel G Gibson, Lei Young, Ray-Yuan Chuang, JCraig Venter, Clyde A Hutchison III, Hamilton 0 Smith.Enzymatic assembly of DNAmolecules up to several hundred kilobases.Nature Methods, ...
Although many SDM methods have been developed, methods that increase efficiency and versatility of this process remain highly desired. Method We present a versatile and simple method to efficiently introduce a variety of mutation schemes using Gibson-assembly but without the need to design uniquely ...