Notebook : August 12

Lab work

A - Aerobic/Anaerobic regulation system

Obtaining Pfnr_RBS-LacZ-Term in PSB1C3

1 - Digestion of Bba_K1155000, Bba_K1155007 and Bba_K1155003

Anaïs, Nadia, XiaoJing

• Pfnr in PSB1C3 (Bba_K1155000) :
  • Buffer FD : 5µL
  • H2O : 38µL
  • Pfnr : 5µL
  • SpeI : 1µL
  • PstI : 1µL

• RBS-LacZ-Term in PSB1C3 (Bba_K1155007) :
  • Buffer FD : 5µL
  • H2O : 23µL
  • RBS-LacZ-Term : 20µL
  • XBal : 1µL
  • PstI : 1µL

• RBS-AmilCP-Term in PSB1C3 (Bba_K1155003) :
  • Buffer FD : 5µL
  • H2O : 33µL
  • RBS-AmilCP-Term : 10µL
  • XBal : 1µL
  • PstI : 1µL

2 - Electrophoresis of digestion pieces

We gonna check if the digestion was good by control the size of digestion pieces.

Well 1 : 6µL DNA Ladder Well 2 : 5µL of digested Bba_K1155000+1µl of 6X loading dy Well 3 : 5µL of digested Bba_K1155007+1µl of 6X loading dy Well 4 : 5µL of digested Bba_K1155003+1µl of 6X loading dy Gel : 0.8%

Expected sizes :

• RBS_LacZ_Term : 3500 kb
• RBS_AmilCP_Term :
• PSB1C3 :

We can't see any band for Bba_K1155000 digestion. We just do it again and let it in the fridge over night. We obtain RBS-LacZ-Term and RBS-AmilCP-Term fragments. the digestion was good. We can now purify it.

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

Objective : obtaining FNR and BphR2 proteins

1 - Electrophoresis of PCR products : RBS-BphR2 part I, BphR2 part I, BphR2 part II, RBS-FNR part I, FNR part I and FNR part II

Damir