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Team:Aberdeen Scotland/notebook/quorumsensing
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Revision as of 15:43, 21 August 2009
University of Aberdeen - Pico Plumber
iGEM 2009
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 (15/06/09)
- Inoculation of transformants - B0015, I732100, R0011 and K081008.
Day 2 Tuesday (16/06/09)
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 (17/06/09)
- 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 (18/06/09)
- 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)
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Day 2 Tuesday (23/06/09)
- Transformation of ccdB resistant e.coli
- using psB3C5, psB3K5, psB3T5, psB4C5, psB4T5 and psB4K5.
- Digestion of purified biobricks - R0011, K093005, R0051, K081008, J37033, R0062 and I732100
- double digest with E + S
- Prepare and run gels using today's digests.
===Day 3 Wednesday (24/06/09)
- Digestion of lysis cassettes- K112022 + K112808
- Using BamHI and BglI
- Repeat digestion of I732100
- Inoculate trasnformed e.coli and grow overnight
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
 Back to Notebook, LacI-Latch Section
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Continue to Notebook, Modeling Section 
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