IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/07/25
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Results from Yesterday
Results Nothing! 1A1 cells were kept in the freedge! B.S. can not be kept in the fridge, low temperatures kill them!
- Yesterday : Single colony of I746001 and I746101 transferred in 10mL of LB + Amp (final concentration 100μg/mL) - Take 8mL of I746001 (AIP sender) and of I746101 - Centrifuge them - Saved 2ml of each in tubes to regrow and add LB - Miniprep I746001 and I746101 pellets (plasmid mniprep kit) : same protocol but 60μL of Zyppy Glution Buffer in step 8 - Restriction Digest : use EcoI and Spel restriction enzymes - Add sequentially : 15μL of SDW + 2μL of 10X Fast Digest Buffer + 2μL of DNA + 1μL of EcoI + 1μL of Spel - Microfuge tubes - Incubate for 10min at 37°C - Heat inactivation of enzymes at 80°C for 5min - GEL 20μLof samples + 4μL of dye
- 10mL of LB + 15μL of 1A1 - Grow at 37°C |