pET-32c

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	The purified product was ligated into pET-32c expression vector and transformed into E. coli BL21 (DE3) pLysS strain using the protocols as given by Sambrook and Russell, 2001.[13]

Protocol tips
The purified product was ligated into pET-32c expression vector and transformed into E. coli BL21 (DE3) pLysS strain using the protocols as given by Sambrook and Russell, 2001.[13]

Publication protocol

Different sets of primers with restriction (RE) sites at their 5' end were designed and synthesised to facilitate directional cloning of PCR products into expression vector. The sequences of these primers with RE sites are: Forward primer (Bam HI and Sal I) 5'-CGC GGA TCC GTC GAC GGC GAA TGG GAT CAG GTA GC-3' and Reverse primer (Hind III and Pst I) 5'-CCC AAG CTT CTG CAG GGG GAA TCC GGT GGG GTT AG-3'. PCR amplification of the gene for rpf-like protein 210 was carried out as described previously. The gel-purified product was digested with Bam HI and Hind III RE enzymes to create cohesive ends for these enzyme sites. The purified product was ligated into pET-32c expression vector and transformed into E. coli BL21 (DE3) pLysS strain using the protocols as given by Sambrook and Russell, 2001.[13]

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Protein Expression Prokaryotic cells - E. coli rpf-like protein using pET-32c from R Verma, Bacteriology and Mycology Division, Mycobacteria Labora.

Manufacturer protocol

Download the product protocol from R Verma, Bacteriology and Mycology Division, Mycobacteria Labora for pET-32c below.

Videos

Check out videos that might be relevant for performing Protein Expression Prokaryotic cells - E. coli rpf-like protein using pET-32c from R Verma, Bacteriology and Mycology Division, Mycobacteria Labora. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.