IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/09/13

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Grow RBS S6
- Put the 10mL of LB with RBS S6 from yesterday into 200mL of LB with antibiotic

- Incubate at 37°C with shaking

RBS W screening
- Plasmid miniprep of RBS W2, 4, 6, 8, 10 9witouh endo wash buffer)

- Nanodrop

- Single digest with EcoRI and XbaI

- Gel1


 * Lane2 : HyperladderI
 * Lane3 : RBS W2 cut with EcoRI
 * Lane4 : RBS W2 cut with XbaI
 * Lane5 : RBS W4 cut with EcoRI
 * Lane6 : RBS W4 cut with XbaI
 * Lane7 : RBS W5 cut with EcoRI
 * Lane8 : RBS W5 cut with XbaI
 * Lane9 : RBS W6 cut with EcoRI
 * Lane10 : RBS W6 cut with XbaI
 * Lane11 : RBS W8 uncut
 * Lane12 : supercoiled ladder

- Gel2


 * Lane2 : HyperladderI
 * Lane3 : RBS W8 cut with EcoRI
 * Lane4 : RBS W8 cut with XbaI
 * Lane5 : RBS W10 cut with EcoRI
 * Lane6 : RBS W10 cut with XbaI
 * Lane7 : RBS W11 cut with EcoRI
 * Lane8 : RBS W11 cut with XbaI
 * Lane9 : RBS W8 uncut
 * Lane10 : supercoiled ladder


 * Result



- W2 with EcoRI : cut

- W2 with XbaI : uncut, self ligation

- W4 with EcoRI : cut

- W4 with XbaI : uncut, self ligation

- W5 with EcoRI : cut

- W5 with XbaI : uncut, self ligation

- W6 with EcoRI : cut

- W6 with XbaI : cut (2 bands), transformation ok for RBS W6

- W8 with EcoRI : impossible to see

- W8 with XbaI : impossible to see

- W10 with EcoRI : cut

- W10 with XbaI : maybe cut, it seems to be 2 bands, but quite difficult to see

- W11 with EcoRI : cut

- W11 with XbaI : uncut, self ligation

Double check of RBS W6 and stock
- Grow RBS W6 in 10mL of LB with antibiotic


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