Team:WPI-Worcester/Harvard-Collaboration

WPI and Harvard Biodesign Collaboration

WPI

We completed the biofilm assay described above with strains of E. coli with modified pili created by the Harvard team. The strains we tested were JW4283, a fim H knockout and negative control for biofilm formation, and C2M+C5 and C9-3+C5, which express modified pili when induced by arabinose and rhamnose. Upon receiving agar stabs from the Harvard team, we streaked plates and incubated them at 37oC for 24 hours. Afterwards, we placed the plates in the refrigerator until we received the arabinose and rhamnose needed to induce pili formation in the experimental strains. Once we had all necessary materials, we grew 5mL liquid cultures of each of the 3 strains in LB in accordance with the protocol. The JW4283 culture was not supplemented with antibiotics or inducers, but the C2M+C5 and C9-3+C5 cultures were supplemented with 5 μL of chloramphenicol, 5 μL of ampicillin, 0.01% arabinose, and 0.5% rhamnose. We prepared the 1:100 dilutions in LB broth and M9 minimal media, which we found to support biofilm formation while designing our biofilm assay. The remainder of the protocol was left unchanged. Below are the results of the assay.
.