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Notebook : August 19

Lab work

A - Aerobic/Anaerobic regulation system

Objective : to obtain Characterize Bba_K1155000, Bba_K1155004, Bba_K155005, Bba_K1155006

1 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term

Nadia

Protocol : EtOH precipitation

We used 20µL of DNA.

Nanodrop :

RBS-LacZ-Term : 38.4ng/µL
RBS-AmilCP-Term : 28.7ng/µL

CONCLUSION

2 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3

XiaoJing

Used quantities :

NirB and RBS-LacZ-Term :
NirB : 3µL
RBS-LacZ-Term : 1µL
Buffer : 2µL
H2O : 13µL
Ligase : 1µL

NirB and RBS-AmilCP-Term :
NirB : 3µL
RBS-AmilCP-Term : 1µL
Buffer : 2µL
H2O : 13µL
Ligase : 1µL

NarG and RBS-LacZ-Term :
NarG : 3µL
RBS-LacZ-Term : 1µL
Buffer : 2µL
H2O : 13µL
Ligase : 1µL

NarG and RBS-AmilCP-Term :
NarG : 3µL
RBS-AmilCP-Term : 1µL
Buffer : 2µL
H2O : 13µL
Ligase : 1µL

NarK and RBS-LacZ-Term :
Nar K : 3µL
RBS-LacZ-Term : 1µL
Buffer : 2µL
H2O : 13µL
Ligase : 1µL

NarK and RBS-AmilCP-Term :
NarK : 3µL
RBS-AmilCP-Term : 1µL
Buffer : 2µL
H2O : 13µL
Ligase : 1µL

3 - Transformation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3 in DH5α

XiaoJing

Protocol : Bacterial transformation

A - Aerobic/Anaerobic regulation system / B - PCB sensor system

Objective : obtaining FNR and BphR2 proteins

1 - Electrophoresis gel of PCR products : BphR2 Part I

Damir

[[]]	Well 1 : 6µL DNA Ladder Well 2 : 40µL BphR2 part I+8µl of 6X loading dye Gel : 1%

Expected size :

BphR2 Part I : 178 kb

We obtain a frangment at the right size. We can purify it.

2 - Gel purification of PCR products : BphR2 Part I