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From 2011.igem.org

Week 6/1/11-6/3/11

Participated in iGEM bootcamp. For the week we we learned bio-basics, CAD basics, Tuberculosis overview, digital logic design and clotho. For clotho we built an application that would show hello world and then we modified the app so that another user could input a part sequence that would be stored in the clotho database. Finally we had a group outing at Jillian's.

Week 6/6/11-6/10/11

On the last day of bootcamp we learned more information on synthetic biology and gave the Wellesley team a tour of the wetlab and our liquid handling robot. The next day I worked with Kyle Jones to look for constitutive promoters in the MIT Registry of Parts. We decided to use BBa_J23100, BBa_J23101, BBa_J23109, and BBa_I14053. The first three are part of the Anderson Collection and are Amp Resistant. These promoters had RBS inside their plasmids. The last constitutive promoter is Kan resistant. Vanessa Yanez and Alberto looked for induced promoters and Shannon and Margueax looked for RBSs. After locating our parts we transformed them and let them sit over night. Unfortunately the parts that we transformed did not work after the plasmid prep. We had low values for the concentration and realized that the cultures that did grow in the LB and amp solution was not E coli. In order to confirm our suspicions we looked at the culture under a light microscope. My cultures seemed to have string like cells which we concluded to be contamination. Only 1 culture worked out of the 18 that were performed that week. Finally we met together to figure out how we wanted to split up the work and what exactly we wanted to do as a team. Margeaux and Alberto did mini prepreps, restriction digest, gel extraction, ligations, and finally transformations for GFP and terminators. The rest of us redid all the old procedures in search for the reason for the contamination.

Week 6/12/11-6/17/11

In order to decrease contamination we autoclaved the pipet tips and used aseptic techniques where applicable. We redid the plasmid preps for the same parts. This time we grew the Ecoli in just LB solution and then in LB with Amp. The next morning we discovered that the cells in the LB and Amp solutions did not grow while the cells in the LB solution did. This proved our suspicion that the plates were our problem. On Monday we prepped the UV inducible promoter for the QiaCube. The UV promoter was the only transformation and plasmid prep that worked. We did the transformation on the existing Amp plates but this time we used a negative control. After incubating the transformed cells we discovered that the negative control did in fact grow colonies. This lead us to making new Amp plates and redoing transformations and plasmid preps for the chosen parts. In the meanwhile, Alberto and Margeaux cut and ligated more GFP and terminator B0015.