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From 2011.igem.org

September 11th - September 17th

Sunday, September 11

~God bless America -- never forget~

Monday, September 12

Afternoon lab work done by: Nancy Li, Charlie Chung

• Prepared ten 3mL LB cultures (with 3µL carbenicillin) of colonies picked from plates dated Sept 3

- RFP + Avi-Tagged pZE12 in DH5α

- vioA + Avi-Tagged pZE12 in DH5α

- vioB + Avi-Tagged pZE12 in DH5α

- vioE + Avi-Tagged pZE12 in DH5α

- RFP + Avi-Tagged pZE12 in MC4100

- vioA + Avi-Tagged pZE12 in MC4100

- vioB + Avi-Tagged pZE12 in MC4100

- vioE + Avi-Tagged pZE12 in MC4100

- Control tube is just a colony with pZE12 (no insert gene)

- LB with carbenicillin (no inoculation with bacteria)

• Incubating at 37°C in the shaker at Weill Hall
• Cultures are to be lysed for release of natively biotinylated RFP and vio enzymes in characterizing streptavidin-coated microfluidics chips
• Did not extensively search for L-tryptophan (starting substrate in the synthesis of prodeoxyviolacein) in the reagents shelf of our Weill Lab space, but does not appear to be in plain view (may be tucked away in an obscure corner?)

Tuesday, September 13

Afternoon lab work done by: Charlie Chung

Objective: Transfer yesterday's 3mL starter culture into today's larger scale culture (20mL LB + 20µL ampicillin)

• Autoclaved two 1L LB at 1:30pm (~1.5 hour cycle)
• Claire's batch of five autoclaved 250mL flasks are ready for use
• Prepared stock solution of 1000x ampicillin (100mg/mL)

- Weigh out 1g ampicillin sodium salt (Sigma)

- Dissolve in 10mL ddH2O

- Syringe filter (0.22µm membrane) the ampicillin solution

• Stored in the bottom shelf of Weill Lab refrigerator (it's in a 15mL conical -- will later aliquot into Eppendorfs)

Evening lab work done by: Claire Paduano

• Prepared cultures of avi-tagged VioA, B, E and RFP in 20mL LB with 20uL ampicillin

Wednesday, September 14

Thursday, September 15

Friday, September 16

Saturday, September 17