Notebook : July 1

summary

For regulator system:

• started the Restriction digestion and ligation to get the BioBrick fnr into plasmid PSB1C3 by using restrictin ensyme EcoRI and PstI.

• Sent an E-mail to M. Nesbeth from UCL for requesting the Biobrick BBa_K239005 which was created by iGEM UCL team in 2009

For PCBs sensor system:

• Designed oligonucleotides for the gene coding for BphR2 (regulator, sensitive for PCBs), Transcriptional regulator of Pseudomonas oleovorans /pseudoalcaligenes group

lab work

• A.aero/anaerobic regulation system
  
  • 1.BioBrick promotor fnr(repressor) in plasmid PSB1C3

Digestion for fnr and PSB1C3

2 enzymes EcoR I and PST I can be used in one common buffer: orange buffer (10X).

For PCR products:

For plasmid PSB1C3:

Ligation

After 3h of digestion, we mixed the digestion products:

Then we performed a precipitation by ethanol in order to inactivate the enzymes. And suspended the deposit by:

The incubation was during 1H30.

• B.PCBs sensor system
  
  • 1.BioBrick BphR2(regulator) in plasmid PSB1C3
    

Using software gene manager to find the oligopeptide for amplification of BphR2.

Notebook : August 23

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K1155000

1 - Digestion of PSB1C3 plasmid and PCR products : Pfnr by EcoRI/ PstI

Zhou

Used quantities :

• PSB1C3 :
  • Plasmid : 4µL
  • EcoRI FD : 0.5µL
  • PstI FD : 0.5µL
  • Buffer FD : 0.8µL
  • H2O : 2.2µL

• Pfnr :
  • Pfnr : 20µL
  • EcoRI FD : 0.75µL
  • PstI FD : 0.75µL
  • Buffer FD : 3µL
  • H2O : 5.5µL