Team:UCLA/Notebook/Protein Cages/14 August 2015
Introdution: An experimental growth period using PCquad Y will be done today.
Procedures:
Colonies successfully grew on the plate that was streaked yesterday. 25mL of LB was added to a culture tube, and 25uL of 100mg/mL carbenicillin was added for a total concentration of 100ug/mL. Using that same tip that aliquotted the antibiotic, a single colony was picked and the entire tip was injected into the tube (tip was a filter tip, but sterility is questionable). The culture tube and plate was then walked over to boelter lab, and incubated in the 37C shake incubator at 10:20AM.
IPTG induction: OD600 was checked periodically. At ~0.6 (0.56 in this case at 4:40PM), 1mM IPTG was added to the culture tube. The culture was grown for another 3 hours before harvesting by centrifugation. Centrifugation was done at 4,500g for 20 minutes. The pellet was resuspended in 1mL of sterile water and centrifuged at max speed for 10 minutes. The total weight of the cell pellet was 0.095g.
Conclusions: Protein purification will be done next week in order to determine if this growth protocol was sufficient.