pET-28b-Cas9-His (BioVector-047327) BioVector NTCC质粒载体菌种细胞基因保藏中心

pET-28b-Cas9-His (BioVector-047327) BioVector NTCC质粒载体菌种细胞基因保藏中心 pET-28b-Cas9-His原核表达Cas9质粒，可用有体外表达和纯化Cas9蛋白，Kan抗性。
For in vitro expression and purification of Cas9 protein
Please see the Schier lab website http://labs.mcb.harvard.edu/schier/ for detailed protocols about target site selection, sgRNA production, stop codon cassette design, Cas9 protein purification, injection and downstream analysis. This plasmid was used to express a His-tagged Cas9 protein in E. coli and purify it by nickel-based affinity purification.The Cas9 protein and sgRNA were incubated at room temperature to form the Cas9 protein/sgRNA complex in vitro, which was then microinjected into zebrafish zygotes. See publication for more details.
pET-28b (Search Vector Database) Backbone size w/o insert (bp) 5368 Vector type Bacterial Expression, CRISPR GROWTH IN BACTERIA Bacterial Resistance(s) Kanamycin Growth Temperature 37°C Growth Strain(s) DH5alpha Growth instructions For expression and purification of Cas9, transform into Rosetta strain. Copy number High Copy GENE/INSERT Gene/Insert name Cas9 Insert Size (bp) 4100 Tag / Fusion Protein His (C terminal on backbone) CLONING INFORMATION Cloning method Restriction Enzyme 5′ cloning site NcoI (not destroyed) 3′ cloning site NotI (not destroyed) 5′ sequencing primer T7
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