IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/07/25

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Results from Yesterday

  • Kan resistance of biobricks
Biobrick Antibiotic Contration (μg/mL) Observations Conclusion
I746001 Kan 25 Many colonies Kan Resistance OK
I746101 Kan 25 Many colonies Kan Resistance OK
Control PUC19 Kan 25 No colony No Kan Resistance


Nothing! 1A1 cells were kept in the freedge! B.S. can not be kept in the fridge, low temperatures kill them!

  • Test if biobrick of last year are really the one we want

- 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

  • Grow 1A1

- 10mL of LB + 15μL of 1A1 - Grow at 37°C