Team:UNIPV-Pavia/Notebook/Week16

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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 22 Week 23 Week 24



Week 16: 09/1/08 - 09/5/08

09/1/08

  • We had to perform these ligations:
    • Lig.22-R0062
    • Lig.31-Lig.22
  • but we didn't have enough Lig.22 and Lig.31 plasmids...
  • We infected 9 ml of LB + Amp with 30 µl of Lig.22 and Lig.31 glycerol stocks.

09/2/08

  • Glycerol stocks/miniprep for Lig.22 and Lig.31.
  • Plasmid digestion for:
    • Lig.22 (E-S)
    • Lig.22 (X-P)
    • Lig.31 (S-P)
    • R0062 (E-X)
  • Run/gel extraction.
  • Ligations:
    • Lig.31-Lig.22 (=Lig.35)
    • Lig.22-R0062 (=Lig.30)
  • We incubated ligations at 16°C overnight.

09/3/08

  • We transformed/plated ligations.

09/4/08

  • Colony PCR for the two ligation plates
Marker 1Kb, Lig.30 (5 colonies), Marker 1Kb, Lig.35 (5 colonies)
  • Gel results:
    • Lig.30 (3rd colony)
    • Lig.35 (gel showed an unexpected contaminant...we decided to re-perform this ligation)

09/5/08

  • Glycerol stock/miniprep for Lig.30.
  • We sent Lig.30 (3rd col) to Primm for sequencing.