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Team:Paris Saclay/Notebook/July/29
From 2014.igem.org
Contents |
Tuesday 29th July
Lab Work
A - The frame
PCR
by Romain
StrainS used: E. coli MG1655Z1 and E. coli MG1655, and oligonucleotides used: FTR-Apra-F and FTR-Apra-R.
Protocol:
- 218µl of H20 MilliQ
- 80µl GoTaq buffer 5X
- 16µl oligonucleotide FTR-Apra-F (10mM)
- 16µl oligonucleotide FTR-Apra-R (10mM)
- 12µl DMSO 10mM
- 32µl MgCl2 (25mM)
- 8µl dNTP (10mM)
- 2µl of GoTaq enzyme
- 2µl of different bacterial cultures in each tubes:
- Tube 1: MG1655Z1 10I
- Tube 2: MG1655 100II
- Tube 3: MG1655 50I
- Tube 4: MG1655 100I
- Tube 5: MG1655Z1 10II
- Tube 6: MG1655 50II
- Tube 7: MG1655Z1 10I positive control with plasmid
Electrophoresis
by Romain
C - Lemon scent
PCR of BBa_K762100
by Sean
Oligonucleotides used: iPS66, iPS67
Oligonucleotides were diluted twice from 100µM to 50µM.
Protocol
Add into a PCR tube the following:
| Component | For a total volume of 50μl |
|---|---|
| H2O | 35.5μl |
| Phusion buffer 5X | 10μl |
| dNTPs | 1μl |
| iPS66 | 1μl |
| iPS67 | 1μl |
| BBa_K762100 | 1μl |
| Phusion DNA polymerase | 0.25μl |
E - Salicylate Inducible Suppressing System
Digestion
by Fabio
BioBrick BBa_J61051 (Salicylate promoter + NahR)
Electrophoresis
by Fabio
Results:
First process to verify the digestion
Second one to separate both parts (only with J61051).
Members there:
- Instructors and advisors: Alice, Solenne and Sylvie.
- Students: Arnaud, Fabio, Romain, Sean and Terry.
