IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/06/25

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Week 1 Competent Cells Put 50mL LB with 1 mL of dH10B (2 500 ml cylinders) in the shaker at 9:23 am. (Vallari Somayaji) OD of the flasks were checked at 1:00 p.m. The OD was found to be about 1.4. This is concentration is too high to make competent cells. Because of time constraints, a sample was transferred to culture tube containing 5 ml of LB. New flasks will be made on 6/26 to try again. Transforming cells The following parts were picked from the iGEM registry plates: Tube Registry Number 1 K654058 - TesA 2 E0240 - GFP Generator 3 J06702 - mCherry 4 R0040 - TetR Promoter 5 R0011 - LacI Promoter 6 B0034 - RBS 7 B0032 - RBS Each part was re-hydrated in 10 μl of water. 30 μl of competent cells, donated from the Haynes lab, was added to each DNA sample. The 7 samples were left on ice for 30 minutes. Heat shocked at 42°C for 35 seconds. Placed back on ice for 2 minutes 750 μl of LB media was added to each tube Tubes were placed in shaking incubator for 1 hour at 32°C 350 μl of supernatant was removed from each tube Pellets were re-suspended and placed onto LB agar plates with chloramphenicol.