Team:Carnegie Mellon/Met-Notebook

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Latest revision as of 03:32, 27 October 2012

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Overview

Lab notebooks were kept using Google docs, and they mostly contain information from the experimental side, including details such as the protocol followed, timestamp of the experiments and the observed results.
Feel free to click on the links below to view the actual lab notebooks and email us if you have any questions!

Notebooks

During the first 2 weeks after final exams, we were mainly focused on settling the logistics of getting the project off the ground. Hence there were no lab experiments and hence no notebook kept.

1. Week 3 was focused transforming two different FAP plasmids we obtained into DH5a and working out the exact construct design

2. Week 4 was focused on testing and comparing the functionality of the two FAP plasmids expressed in BL21DE3 cells using two variants of Malachite Green (5% EtOH and PBS)

3. Week 5 was focused on cloning our synthesized Spinach cassette into our vector, and conducting dosage and temporal curves using the FAP and MG(5% EtOH) used.

4. Week 6 was focused on Spinach. We ran a gel and sequenced our Spinach transformant and also PCR amplified the original cassette to obtain more copies.

5. Week 7 was focused on Spinach cloning. We tried another round of cloning of Spinach into the FAP vector and a pET vector. Also provides the basis for the first T7 promoter designs.

6. Week 8 was focused again on cloning Spinach into the FAP vector and the pET vector after another negative result of cloning. Finalized the T7 promoter designs

7. Week 9 was focused on characterizing the Spinach cassette in a pET vector. Efforts continued to create a clone with both the FAP and the Spinach Cassettes.

8. Week 10 focused on cloning Spinach and the FAP into a different vector with entirely new restriction sites. Began preparation for a cloning round in URA- yeast as a backup plan. Cloned cassettes into pIVEX vector.

9. Week 11 included cloning round in URA- yeast using homologous recombination. Characterized Spinach using a functional assay in pIVEX vector.

10. Week 12 is a brief document detailing the PCR reaction, which is in preparation for the Fall laboratory rounds or cloning.

11. Fall notebook is a consolidated log of all our experiments carried through Fall, including the final characterization and cloning experiments.

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