"
EPF-Lausanne/8 July 2009
From 2009.igem.org
Wet Lab
1. R. Palustris culture grew. A glycerol stock has been done. A pellet is on the fridge level 2, waiting for a miniprep.
2. iGEM parts have been transformed:
| Part | Characteristic | Resistance | Well (Kit Plate) |
|---|---|---|---|
|
Terminator |
A |
13D (1) | |
|
Promoter LacI |
A |
1D (1) | |
|
RBS |
A |
1H (1) | |
|
RBS-GFP-TER |
A |
12M (1) | |
|
RBS-mRFP-TER |
A |
22O (1) | |
|
pTetR-RBS |
A |
13B (1) |
Cloning Strategy
Inducible LOVTAP biobrick strategy
- Problem to overcome:
- PstI sites in LOVTAP sequence.
- Our goal:
- Biobrick consisted of LacI promoter-RBS-LOVTAP-Term (in this order).
- Material:
- Biobrick of LacI promoter, RBS, LOVTAP, Term separately ( LOVTAP obtained from previous section and the rest from iGEM Spring 2009 distribution Kit plate).
- Strategy:
- LacI promoter-RBS ligation with iGEM protocol (LacI promoter digested with ES, RBS digested with XP, plasmid containing an other antibiotic digested with EP).
- LOVTAP is digested with ES and inserted into Term plasmid (which was digested with EX previously).
- Finally, LacI promoter-RBS digested with ES to be inserted into LOVTAP-Term plasmid, digested with EX.
People in the lab
- Heidi, Tu, Nath, Rafael, Basile
