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Team:Grinnell/Notebook
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==Notebook== | ==Notebook== | ||
Revision as of 00:25, 11 June 2011
Contents |
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
- Figure 1: 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. 
- Figure 2: Gel results for
transformation of ligations. Lane 1:
ladder; Lane 2: digested plasmid
with rsaA; Lane 3: digested plasmid
with rsaA and esp; Lanes 4-8:
digested plasmids from various
colonies with esp.
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 pSB1C3 with EcoRI and PstI. We then ligated esp, rsaA, and esp and rsaA to pSB1C3 and transformed into E. coli Top10. (Protocol)
Inoculated liquid cultures with colonies from transformations.
Performed digestion of results from transformation and ran on gel (figure 2).
Transformed BBa_K081005, a BioBrick part containing a gene for constitutive promotor and RBS, into E. coli Top10.
