Team:UNIPV-Pavia/Notebook/Week9

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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 8: 07/14/08 - 07/17/08

07/14/08

  • We transformed 1 µl of the 6 ligations stored at -20°C. We plated transformed bacteria and incubated plates at 37°C overnight.
  • Plasmid digestion for:
BBa_R0079 (E-X) BBa_B0030-BBa_C0078-BBa_B1006 (E-S)
  • Gel run/cut/gel extraction for the 2 parts.
  • Ligation: BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079.
  • We incubated ligation at 16°C overnight.



07/15/08

  • We transformed 1 µl of BBa_B0030-BBa_C0078-BBa_B1006-BBa_R0079 ligation and plated transformed bacteria. We incubated the plate at 37°C overnight.
  • All the 6 overnight plates showed colonies. We performed colony PCR for (we decided to pick up more colonies for the ligations with long inserts):
  1. BBa_J23100-BBa_B0030 (S-P) -BBa_C0012 (standard ligation) - 10 colonies
  2. pSB1AK3-BBa_J23100-BBa_B0030-BBa_C0012 (double ligation) - 7 colonies
  3. BBa_J23100-BBa_B0030-BBa_C0040-BBa_B1006-BBa_R0040 - 3 colonies
  4. BBa_R0051-BBa_B0030-BBa_C0062-BBa_B1006-BBa_R0062 - 3 colonies
  5. BBa_B0030-BBa_C0051-BBa_B0030-BBa_C0079-BBa_B1006 - 10 colonies
  6. BBa_B0030-BBa_I15009-BBa_B1006 - 3 colonies
Picked up colonies in LB + Amp