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From 2010.igem.org

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Contents 1 6/14-18/10 (Boot Camp) 2 6/22/10 3 6/23/10 4 6/24/10 5 6/25/10 6 6/28/10 7 6/29/10 8 6/30/10 9 7/1/10 10 7/2/10

6/14-18/10 (Boot Camp)

DNA extracted from kit

Part: [http://partsregistry.org/partsdb/get_part.cgi?part=BBa_I746200]

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6/22/10

We poured our Kan, Amp, Tet, Kan+Amp, Kan+Tet, Amp+Tet antibiotic agar plates.

Antibiotic Concentrations

• Amp: 50mg/500ml

• Kan: 31.97mg/500ml

• Tet: 6mg/500ml

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6/23/10

Resuspended 12O and 6G DNA from the iGEM Kit and transformed them into our competent cells.

6G = r0010(inducible promoter) 12O = e0840(rbs30-gfp-2xterm)

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6/24/10

Ran a gel of our PCR product to make sure that our taq polymerase master mix was working.

Selected single 12O and 6G colonies and transferred them to LB and incubated for 18 hours.

Let 1 12O and 1 6G plate incubate at RT. Let 1 12O and 1 6G plate incubate at 37°C. Intended to observe lawn growth.

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6/25/10

Low DNA yield; Plan to redo DNA extraction from Kit

Found pink colonies in 12O plates. Determined to be contaminations. Plates were thrown out.

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6/28/10

Kit to Stock

1-12O (r0010: inducible promoter)

1-6G (e0840: rbs30-gfp-2xterm)

3-20M (K124017: Bacteriophage Lysis Cassette S105, R, and Rz)

Plates were incubated at 37°C overnight (starting at 2:30 pm).

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6/29/10

Selected single 1-12O, 1-6G, 3-20M colonies and grew them in Amp-LB broth for 18 hours (starting at 4:15 pm).

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6/30/10

2 Sleeves of K and A plates respectively

Meeting with Ruffin (BIF)

Need to talk to: Aaron Packman, Wei Zhang, Kimberly Grey

Most likely going to use water immersion Leica (Confocal)

Need to email Ruffin with additional data (biofilm thickness is about 5~50um)

miniprepped 112o,320m,16g: mostly below 20ng, 1 above 100ng; stored in -20

reminiprepped after growing in liquid LB for 3 hours

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7/1/10

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7/2/10

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