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Team:Michigan/Quorum Sensing
From 2010.igem.org
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~1.5 hrs work | ~1.5 hrs work | ||
| + | Created protocol/plan for experiment. [[Testing AI-2 Response]] | ||
--[[User:Infekt|Infekt]] 00:26, 18 July 2010 (UTC) | --[[User:Infekt|Infekt]] 00:26, 18 July 2010 (UTC) | ||
Revision as of 00:28, 18 July 2010
| Sunday | Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | |
| Week 1 | - | - | - | - | - | - | - |
| Week 2 | - | - | - | 7/7/2010 | 7/8/2010 | - | - |
| Week 3 | - | - | - | - | - | - | - |
Quorum Sensing Team
Members include Alex Pyden, Marcus Lehr, Jennifer Hong, Eric Raynal, Katie Miskovich, and Audra Williams
7/7/10
Jennifer and Alex - in Lin Lab
Made CaCl2 stock solution: 0.1M - 50mL
- molar mass = 111 g/mol
- (0.1 mol/L)(.05 L)(111 g/mol) = 555mg
- dissolved 555 mg CaCl2 in 50mL DIwater
- vacuum filtered into 50mL tube
Made ampicillin stock solution:
- 1 g ampicillin
- 5mL DIwater
- ethanol
- filtered by syringe into 15mL tube
- put into ten 1mL alliquots in 1.5mL tubes
- stored in -20°C in ERB lab
Inoculated E. coli DH5α from frozen stock into 12 mL LB broth
- put in 30°C, 200 rpm shaking, overnight
~1.5 hrs
7/8/10
Alex, Jennifer and Eric - in ERB
Made LB agar plates w/ ampicillin
- 10 g LB broth
- 7.5 g agar
- 500 mL DI water
- autoclaved mixture at 121°C for 30 min (sterilize time), following Mike Nelson's protocol
Created protocols for Obtaining Deionized Water in the ERB and ERB Spectrophotometer
- uploaded to Team:Michigan/Protocols section
poured 20 LB-amp plates (large)
- left to solidify in ERB 1230
- 12 plates were used by Ann for biobrick transformations
- 8 plates were stored in 4°C ERB
~3.5 hrs work
Ann, Marc, Audra and Katie - in Lin Lab
Performed biobrick transformations using heat shock
- Followed protocol for Transformation-heat shock
- protocol located under DNA Manipulation in Team:Michigan/Protocols section
After second washing:
- ODs for sample 1 and 2 were 0.378 and 0.358 respectively
4 hrs of work
7/17/10
Alex & Jennifer - in ERB
Yesterday, Marcus stored newly obtained strains in 4°C in ERB
- W3110 w/ plasmids pTC6 & pET-GFP
- AI-2 reporter (E. coli, amp and kan-resistant)
- MDAI2 w/ plasmids pCT6 & pET-GFP
- AI-2 reporter & LuxS null mutant (E. coli, does not produce AI-2, amp and kan-resistant)
Removed these strains from 4°C -- they are stored in soft agar stab cultures Made one streak plate on LB+amp for each strain from stabs
- placed in 374°C (rm. 1239)
Made one 2mL LB+amp broth cultures in a 15mL tube for each strain from stabs
- placed in 30°C, 200 rpm shaking (rm. 1230)
Placed agar stabs and LB+amp broth in 4°C
~1.5 hrs work
Created protocol/plan for experiment. Testing AI-2 Response
--Infekt 00:26, 18 July 2010 (UTC)
