Team:Aberdeen Scotland/notebook/quorumsensing

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University of Aberdeen iGEM 2009


Contents

Quorum Sensing Notebook

Week 1 - Research and Biobrick Rescues (08/06/09 - 12/06/09)



Day 1 Monday (08/06/09)

  • Researching the Registry for Biobricks
    • Identified K081008, J23100, C0062, K091156, B0034,B0030, B0015, R0061 and R0062 for possible use in our project.
    • Other biobricks identified for other sub-projects include R0011.



Day 2 Tuesday (09/06/09)

  • Researching the literature for information on Quorum Sensing.
  • Researching for information on LVA tags



Day 3 Wednesday (10/06/09)

  • Preparing for biobrick rescue and transformation
    • Prepare LB and Agar
    • Prepare Cacl2 solution,
    • Prepare 1M MgCl2, 1MMgSO4 and 2M glucose solutions (for SOC medium)
    • Inoculate e.coli and grow overnight



Day 4 Thursday (11/06/09)

  • Rescue Biobricks (K081008, J37033, B0015, J23100, R0011, R0062, K112808, C0051, C0040, I732100, R0051, K145001 and B0034)
  • Prepare competent e.coli, using calcium chloride method
  • Transform e.coli with biobricks listed above
    • grow overnight on agar/amp plates.



Day 5 Friday (12/06/09)

  • Results from yesterday indicate that only I732100 was successfully transformed into e.coli
  • Identify possible recipient plasmids for QS module (psB4C5).
  • Propose a cloning strategy
  • Team meeting



Week 2 (15/06/09 - 19/06/09)



Day 1 Monday (08/06/09)

  • Inoculation of transformants - B0015, I732100, R0011 and K081008.



Day 2 Tuesday (09/06/09)

Inspection of inoculations, after overnight growth, revealed, as suspected, that only I732100 had grown

  • Perform plasmid prep on I732100
  • Digest I732100
    • 1 Double digest E+ S
    • 4 single digests - E, S, X and P
  • Prepare TSS buffer, for TSS method of competency (to be performed tomorrow)
  • Inoculate e.coli and grow overnight for tomorrow's TSS method.



Day 3 Wednesday (10/06/09)

  • Rescue additional biobricks - K112022, R0040, I732094, K093005, E0840
  • Prepare competent e.coli using TSS method.
  • Transform with biobricks rescued today + B0015, R0011, R0051, K112808 and K081008, grow overnight



Day 4 Thursday (11/06/09)

  • Prepare for miniprep
    • inoculate transformed e.coli and grow overnight in liquid medium
  • Create master plates to keep record of colonies used



Day 5 Friday (12/06/09)

  • Perform miniprep to purify R0011, R0051, B0015, K112808, K081008
  • Team meeting



Week 3 22/06/09 - 26/06/09



Day 1 Monday (22/06/09)

  • Transformation of ccdB resistant e.coli
    • using psB3C5, psB3K5, psB3T5, psB4C5, psB4T5 and psB4K5.
  • Digestion of purified biobricks -



Day 2 Tuesday (09/06/09)

Week 4 29/06/09 - 03/07/09

Week 5 05/07/09 - 10/07/09

Week 6 13/07/09 - 16/07/09

Week 7 20/07/09 - 24/07/09

Week 8 27/07/09 - 31/07/09

Week 9 03/08/09 - 07/08/09

Week 10 10/08/09 - 14/08/09

Week 11 17/08/09 - 21/08/09

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