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.

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

Protocol : hight copy plamid ectraction

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.