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From 2012.igem.org

week 2: 7 may - 11 may

office work

lab work

7 may

E. coli in liquid LB

3 plates per strain were picked and grown in 25 ml LB. all plates grown well accept for amp plates, due to drying effects.

8 may

10:00 Continue growth. All 3 strains in shaker, 180 RPM, 30 degree celcius.

4 x 1 L SOB is made

16:55 culture mach1 and DH5X are plated 10x and 1000x diluted on LB plates. Dilutions are done with demi water.

Plates were placed in 37 degree Celcius stove

9 may

9:15 10x dilution was to low, overgrown 1000x was nearly overrown, but still acceptable.

picked 1 MACH1 and 1 DH5X colony and grown in 10 ml SOB medium, 37 degree Celcius, 180 RPM

10 may

MACH1 competent cells followin pinky's protocol. with the following diviations

• 4x 500 ml SOB
• SLA 3000 rotor

OD harvest at: 0.5 -0.45

11 may

CCMV protocol inoculated at 8:45 IPTG at 11:45

centrifugation in greiner tubes

pellet stored at -20 degree Celcius, 50 ml greiner tubes

Miniprep following the Gene Jet Fermentas protocol

• pet28
• standard 10 stock 1
• standard 10 stock 7