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Revision as of 00:29, 18 July 2010 (view source) Infekt (Talk | contribs) (→7/17/10) ← Older edit		Revision as of 23:32, 19 July 2010 (view source) Infekt (Talk | contribs) (→Quorum Sensing Team) Newer edit →
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	--[[User:Infekt|Infekt]] 00:26, 18 July 2010 (UTC)		--[[User:Infekt|Infekt]] 00:26, 18 July 2010 (UTC)
		+
		+	== 7/19/10 ==
		+	''Alex and Marcus''
		+
		+	7/17 plates are no good -- they needed kanamycin
		+	*plates were discarded
		+	*broth cultures had kanamycin added a day later -- too late to be cryostored, but they were transfered to ERB 4°C
		+	Made LB agar

---

Revision as of 23:32, 19 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)

7/19/10

Alex and Marcus

7/17 plates are no good -- they needed kanamycin

• plates were discarded
• broth cultures had kanamycin added a day later -- too late to be cryostored, but they were transfered to ERB 4°C

Made LB agar