Team:Evry/Notebook/Sensing/PCBs/08-28-2014

Sensor construction bphR2/PbpR1

PCR colony has been done with colonies of the transformation bphR2 in pSB1C3. The tail expected was 1242bp and we obtained it:


A nanodrop has been done on the colony 1 (48ng/µl) and colony 2 (4&,4ng/µl) and the colony has been send to sequencing


With PCR clean up of each part, golden gate has been done. Volumes for each part has been calculated with cloning bench application.
Mix DNA contains:

• J23114-RBS (82,1ng/µl):0,06 µl
• bphR2 (139,3 ng/µl) : 0,327µl
• bphR1 (101,1ng/µl): 0,21µl
• RFP (75,3 ng/µl): 0,46µl
• Terminator (28,1ng/µl) : 0,37µl
• pSB1C3,G3 (107,2ng/µl) : 0,93µl
  
  Mix enzyme contains:
  • 1,5µl 10X T4 ligase buffer
  • 0,5µl BSAI
  • 0,5µl T4 ligase
  • 2,5µl
  
  Aug 28