Contents 1 Notebook : July 10 1.1 Summary: 1.2 Lab work 2 Notebook : August 23 2.1 Lab work 2.1.1 A - Aerobic/Anaerobic regulation system 2.1.1.1 Objective : obtaining Bba_K1155003, Bba_K1155007 2.1.1.2 1 - Transformation of Bba_I732019 and Bba_B0010 in DH5α 2.1.1.3 Objective : obtaining biobricks in PSB3K3 2.1.1.4 1 - 2.1.2 B - PCB sensor system 2.1.2.1 Objective : obtaining Bba_K1155001, Bba_K1155002, BphR2 protein 2.1.2.2 1 - Transformation of ligation of PSB1C3 and BphA1 or BphR1 or BphR2

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.