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The BglB gene was codon-optimized for E. coli and cloned into a pET29b+ vector using Gibson assembly. Site-directed mutagenesis, utilizing the Kunkel method was used to generate mutations to BglB via the Transcriptic cloud laboratory platform.   BglB mutant plasmids and E. Coli cells  were incubated together  on ice, allowing the plasmid to permeate mix with  the E. coli's surroundings. Heat shock at 42 degrees C and immediate incubation on ice result resulted  in E. Coli's uptake of exogenous genetic material. The addition of terrific broth and incubation at 37 degrees C for an hour assist in the recovery of E. Coli and the incorporation of the plasmid. The recovered cells were spread evenly on LB agar plates with kanamycin resistance and were incubated at 37 degrees C overnight. Multiple colonies on the agar plates should be visible within 12 hours. Growth media, containing terrific broth and kanamycin solution, was created. A single colony was scraped from the incubated LB agar plates and dipped into the growth media in a falcon tube. The single colony contains the mutant plasmid, which contains a gene resistant to kanamycin. In the growth media, the transforming plasmid survives while the other bacteria are susceptible to this antibiotic. This process ensures that only transformed cells are able to grow. The falcon tube was covered and incubated with shaking for 37 degrees C for 24 hours.