Notebook : July 10

Summary:

For regulation system:

• prepared the solution of BioBrick fnr repressor in PsB1C3 plasmid for DNA sequencing.
• the terminator transformation of BBa_B0010 did not work yesterday, a second transformation had been done for it.
• The transformation for RBS+LacZ+terminator plasmid into competent cells was performed.
• after the transformation PSB3K3 plasmid in competent cells, these cells were cultured in a liquid nutritive medium.

For PSBs sensor system:

• the ligation products were transformed into competent cells and were cultured on solid medium with their specific antibiotics.

Lab work

A.aero/anaerobic regulation system:

• BioBrick RBS+LacZ+terminator in plasmid PSB1C3
• BioBrick RBS+amilCP+terminator in plasmid PSB1C3

Transformation

Transformation for BBa_B0010 dit work, we observed 0 colonies on the Petri dish, We decided to redo another transformation

Transformation for BBa_I732019(RBS+LacZ+terminator BBa_B0010):

the construction of RBS+LacZ+terminator BBa_B0010 is already done and stocked in iGEM BioBrick bank named BBa_I732019 12G p4 kit 2012. So we just suspended the BioBrick with 10µl water, transformed them into 100µl competent cell.

Cloning for plasmid PSB3K3:

results of transformation and cloning: 34 colonies grown.

We picked up 2 colonies, seeded them in liquid medium(LB+chloramphenicol), incubation at 37°C, 200rpm during one night.

• Contruction for BioBrick promoter BphR1, BphR2, BphA1

From the ligation products which we performed yesterday, We inserted those BioBrick into competent cell( 5µl of ligation product with 50µl competent cell). Then cloning on Petri dish(LB with antibiotics chloamphénicol), incubation during 1 night at 37°C.