Plasmids 101: Common Lab E. coli Strains - Addgene
网页The majority of all common, commercial lab strains of E. coli used today are descended from two individual isolates, the K-12 strain and the B strain. K-12 was isolated from a patient in 1920 and eventually led to the common lab strains MG1655 and its derivatives DH5alpha and DH10b (also known as TOP10).
High rates of plasmid cotransformation in E. coli overturn the ...
网页2022年7月7日 — Using low concentrations of several plasmids that encode different fluorescent proteins, under the same selective antibiotic, we show that E. coli bacteria readily accept multiple plasmids ...
Addgene: Protocol - Bacterial Transformation
网页If you used 100-1000 ng of total DNA in a ligation you will often get more colonies if you use 1 μl of a 1:5 or 1:10 dilution rather than 1 μl directly. Learn how to transform E. coli with your plasmid of interest.
Bacterial Transformation Workflow | Thermo Fisher Scientific - US
网页E. coli is the most common bacterial species used in the transformation step of a cloning workflow. In its natural state, the competency of E. coli is very low (10-5 −10-10) [1] thus cells must be made competent for efficient transformation. The protocols for preparing competent cells vary by choice of transformation method—heat shock or ...
In vivo cloning of up to 16 kb plasmids in E. coli is as simple
网页2017年8月24日 — Here, we describe a simple, fast, and economical cloning method based on RecA- and RecET-independent in vivo recombination of DNA fragments with overlapping ends using E. coli. All DNA fragments were prepared by a 2-consecutive PCR procedure with Q5 DNA polymerase and used directly for transformation resulting in 95% cloning accuracy and zero ...
Fast and antibiotic free genome integration into - Nature
网页2020年10月5日 — Genome-based Escherichia coli expression systems are superior to conventional plasmid-based systems as the metabolic load triggered by recombinant compounds is significantly reduced. The...
Direct and convenient measurement of plasmid stability in lab …
网页2017年7月6日 — We also performed direct stability studies of a currently circulating resistance plasmid in a clinical isolate, strain EC958, which is a member of the rapidly expanding global ST131 E. coli...
E. coli Plasmid Vectors: Methods and Applications - SpringerLink
网页A comprehensive collection of readily reproducible techniques for the manipulation of recombinant plasmids using the bacterial host E. coli. The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones.
Plasmids for Controlled and Tunable High-Level Expression in E. coli
网页2022年11月7日 — In E. coli, the pACYC backbone has a low plasmid copy number, pCOLA and pCDF have medium plasmid copy numbers, and the pET backbone has a high plasmid copy number (2, 29) (Fig. 1A). Plasmid copy number is often directly related to gene expression because more DNA templates are available for transcription, though this relationship is not always ...
Recombinant protein expression in Escherichia coli: advances and ...
网页2014年4月17日 — Plasmid multimer formation, an important cause of instability, relies on the recombination system of E. coli (Summers et al., 1993). All three strains have their λDE3-containing derivative (available at Novagen) so the T7 RNAP system can be used.
- 某些结果已被删除