Notebook : August 9

Lab work

A - Aerobic/Anaerobic regulation system

1 - Obtaining Δ fnr E. coli strain by transduction to test our biobricks

Abdou, Anais, Damir, Nadia, XiaoJing

We do the exprience again because the lysis made on wensday didn't happen. It's probably because the phage strain was too old ( 2001)

Protocol : transduction

Picture: lysed cells comparison. 0µl phage(control): the petri dish is cloudy, bacteria are not lysed. 50µl phage: the petri dish is clear, bacteria are lysed by phages. We used a wild type strain to keep a stock and a Δ fnr E. coli strain to obtain phages that might have encapsidated a Δfnr::Km fragment.

10µl,50µl and 100µl petri dishes are clear so phages are multiplied.

We let the antibiotic over night to select the right strain.

2 - Obtaining RBS_lacZ_ter.

Abdou

Clone 10,14 and 15 plamid extraction using nucleospin kit.

Protocol : hight copy plamid extraction

Results: concentration measured by nanodrop

• clone 10: C=38ng/µl 260/280= 1.78
• clone 14: C=48.5ng/µl 260/280=1.90
• clone 15: C=52 ng/µl 260/280=1.78

We still have to sequence plasmids in order to verify our results.

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

Objective : Obtaining FNR and BphR2 proteins (Gibson assembly)

1 - Gibson assembly.

August 1st PCR purification to be sure about the experiment. BphR2 part1, BphR2 part2 and RBS_BphR2_part1, FNR part1, FNR part2 and RBS_FNR_part1.

Protocol : PCR_clean_up

Results:

we have no fragment so we must do again these PCRs