总之，Golden Gate克隆法是一种强大的DNA重组工具，特别适用于多片段组装的DNA分子，但在使用之前需要考虑其特定的优势和劣势，并根据研究需求做出选择。参考文献：Engler, Carola et al. “A one pot, one step, precision cloning method with high throughput capability.” PloS one vol. 3,11 (2008): e364...
总之,Golden Gate克隆法是一种强大的DNA重组工具,特别适用于多片段组装的DNA分子,但在使用之前需要考虑其特定的优势和劣势,并根据研究需求做出选择。 参考文献: Engler, Carola et al. “A one pot, one step, precision cloning method with high throughput capability.” PloS one vol. 3,11 (2008): e3647...
使用Gateway 克隆系统会产生含有一个编码8个氨基酸的attB重组痕迹的结构,但是Golden Gate连接能设计成无痕的,并且比许多商业化的克隆方法都要便宜。 02Golden Gate和合成生物学 合成生物学已经将Golden Gate克隆方法用在了分子克隆策略中,有时称其为MoClo,这个策略利用了IIS型限制酶BsaI和BpiI/BbsI可一次性有效组装多达...
Addgene 提供两种基于 Golden Gate 的 gRNA 组装方法,可让您高效地将多达 7 个 gRNA 克隆到一个目标载体中,从而轻松实现多重化。 金门克隆的缺点 Golden Gate并非100%不依赖于序列,为避免预料之外的酶切,IIS型限制酶识别位点不能存在于所连接的片段内部。解决这个问题的方法是改造目的片段,通过PCR扩增在片段内识...
One of these methods, Golden Gate cloning, allows assembling up to nine fragments at a time in a recipient plasmid. Cloning is performed by pipetting in a single tube all plasmid donors, the recipient vector, a type restriction enzyme and ligase, and incubating the mix in a thermal cycler....
A one pot, one step, precision cloning method with high throughput capability和Transcriptional activators of human genes with programmable DNA-specificity和sembly of custom TALE-type DNA binding domains by modular cloning和Efficient design and assembly of custom TALEN and other TAL effector-...
The continual demand for specialized molecular cloning techniques that suit a broad range of applications has driven the development of many different cloning strategies. One method that has gained significant traction is Golden Gate assembly, which achieves hierarchical assembly of DNA parts by utilizing...
Learn the principles of Golden Gate cloning and how to simulate standard or batch Golden Gate cloning.
The Golden Gate cloning technique has been proven to be an efficient tool for a variety of cloning setups. The utilization of restriction enzymes which cut outside of their recognition domain allows the assembly of multiple gene fragments obtained by PCR amplification without altering the open ...
另外,为了帮助大家更好的理解Golden Gate克隆技术的原理,小远专门翻译了一条视频帮助大家理解,大家一定要点击文末彩蛋观看视频哦~ References: Engler C, Kandzia R, Marillonnet S. A one pot, one step, precision cloning method with high throughput capability. PLoS One. 2008, 3(11): e3647....