Team:UNIPV-Pavia/Notebook/Week15

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Notebook



Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7
Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14
Week 15 Week 16 Week 17 Week 18 Week 19 Week 20 Week 21



Week 15: 08/25/08 - 08/29/08

08/25/08

  • Pladmid digestion for:
    • R0051 (S-P)
    • R0040 (S-P)
    • B0030-C0061-B1006-R0062-B0030-E0040-B1006 (E-X) (=Lig.b (E-X))
    • B1006 (E-X)
    • Lig.12 (E-S)
  • Run/gel extraction.
  • Ligations:
    • BBa_R0051 (S-P) - BBa_E0240 (X-P) (for promoter test)
    • BBa_R0040 (S-P) - BBa_E0240 (X-P) (for promoter test)
    • Lig.12 (E-S) - B0030-C0061-B1006-R0062-B0030-E0040-B1006 (E-X) (for AND logic gate test)
    • Lig.12 (E-S) - BBa_B1006 (E-X) (to re-perform mutated assemblies)
  • We incubated ligations at 16°C overnight.
  • We ordered 3OC6HSL (Sigma).

08/26/08

  • We transformed/plated ligations.

08/27/08

  • Plates grew correctly. We checked colony fluorescence of three plates under UV rays:
    • R0051-E0240 glowed
    • R0040-E0240 glowed
    • Lig.12-Lig.b (R0051-B0030-C0062-B0030-C0061-B1006-R0062-B0030-E0040-B1006) glowed
  • Colony PCR for 5 colonies of Lig.12-B1006 (called Lig.22).
Lig.22 (5 colonies), Marker 1Kb
  • Gel results: we chose 1st colony to grow a 9 ml overnight culture.

08/28/08

  • Glycerol stocks/miniprep for Lig.22.
  • We sent purified plasmids to Primm for sequencing.
  • We infected 9 ml of LB + Amp with 30 µl of R0062 glycerol stock.

08/29/08

  • Glycerol stock/miniprep for R0062.