Technique: Probe Phage Genomes for Host Binding Proteins
Type
Blog Post
...DNA fragments in E. coli
Now that random DNA fragments have been cloned into E. coli cells, isolated colonies...fragments so that these fragments can be cloned into E. coli and subsequent proteins screened for cell-binding...vector designed for protein overexpression in E. coli, such as pET30a. If working with DNA that is refractory...resultant plasmid libraries into chemically competent E. coli and analyze transformants for appropriately sized...colonies. Transform the resultant plasmid DNA into E. coli BL21 (DE3) cells (these have the T7 polymerase and...membranes for 15 min to kill any remaining live E. coli. At this point, it can really help to incubate the...conditions. Note: if using fast-growing bacteria like E. coli or Salmonella, or any cells that bind strongly to...