840:153g:Projects/project2/2008/09/11

The sweet smell of ...E.coli?

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Project presentation & lab activity planning Our first meeting today was at 8.00am at the library for discussion about our presentation on our project later in class. Some of us met about 1.30 pm to discuss more about the project and the presentation. During class: Corey represented the team and gave Axel and the other members of the class an overview and some details about the project. There were not very many questions except that we have to make sure that the E.coli strain that we are proposing to use actually exist in the registry and if not can we obtain the strain from the initial investigators that work with that particular strain as it is very essential that we obtain a strain that is EnvZ deficient in nature. Once the presentation was done, we moved on to the lab to plan out our project and the list of steps that needs to be carried out for our project starting from next Tuesday. The following is the outcome of our discussion and the decision that was made: Aaron and Matt will be obtaining details about the E.coli strain that we are interested in and mail the team that could provide us with that particular strain of E.coli The initial step of our project is to obtain the Biobricks listed in our proposal from the registry. These bio bricks need to be ligated into vectors and transformed in competent DH5alpha cells for long term storage and for large amounts of DNA. The first step is to prepare all necessary lab essentials before starting the project as well as preparing competent cells. The vector that will be used is pBlueScripT which will be provided by Axel. Since Corey has made competent cells before, he will be in charge of making it and the other members of the group will prepare all necessary solution on tuesday. Surabhi and Diveena: Prepare LB Broth & LB+ Agar(For plates that will be needed on Thursday to streak transformed cells) Corey and Aaron: TSS Buffer Matt and Jared: 0.1M CaCl2 solution Sterilizing of tips, tubes, water and spatulas will also be done. We also fomulated a list of initials for everyone in the group to be used throughout the project: Corey: CH Matt: MS Surabhi: SG Diveena: DV Jared: JK Aaron: AM The following is a list of items that we need to start working in the lab on tuesday: 1.6mL & 0.6mL tubes 4 autoclavable 500mL bottles DH5 alpha cells Petri dishes 70% ethanol Stir bars Beakers & Conical flasks Gloves Trays for use in the autoclave CaCl2 Agar Spatulas Weight boats Balances Autoclaved indicator tapes pH meter 0.1M HCl & 0.1M NaOH Markers Aluminum Foil Pipettes LB powder Measuring cylinders Updated by Diveena