Team:Grinnell/Notebook

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Revision as of 20:29, 10 June 2011

Notebook

Week 1

PCR primers designed according to Biobrick specifications and ordered for rsaA C-terminal and WT esp. Because of the discrepancy in GC content between Staphylococcus epidermidis and Caulobacter, we also optimized the esp sequence for Caulobacter for later synthesis with a higher resulting GC content.

Prepared competent cells of E. coli Top10. (Protocol)

Streaked plates of Caulobacter, Staphylococcus aureus, and S. epidermidis.

Template DNA for PCR prepared from colonies using [http://www.bioventures.com/products.php?cid=10005| GeneReleaser]. (Protocol)

Week 2

• 
• PCR products on DNA gel.
  Lane 1: ladder; Lane 2: rsaA from
  liquid culture Caulobacter; Lane 3:
  rsaA from plate culture Caulobacter;
  Lane 4: esp from S. epidermidis.

Performed a transformation experiment (protocol) to test the efficiency of our competent cells make in Week 1. Results were successful with about a 0.7 x 10^7 CFU/μg of pUC19.

Purified PCR product (protocol) and digested esp with EcoRI, SpeI, and PstI; rsaA with EcoRI, XbaI, and PstI; and spB1C3 with EcoRI and PstI. We then ligated esp, rsaA, and esp and rsaA to spB1C3 and transformed into E. coli Top10. (Protocol)

Inoculated liquid cultures with colonies from transformations.