Team:TU-Delft/Notebook/2013/08/29/
From 2013.igem.org
Notebook
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29th of August
Lab work
1. Transformation of pTET:cI:TT was successful. Inoculated it on LB broth.
2. Made glycerol stocks of SUMO+Peptides and pT7:Maximin:His6. Mini prep was also carried out on all. They were sent for sequencing.
3. Transformation of pT7, BBa_I712074, was done on XL1 Blue cells.
4. Restriction digestion of pTET:cI:TT with E+S was run on gel:
5. 1/50 dilution of the pT7:Maximin H5:His6 inoculation was done at 11:00 am.
6. At 2:00 pm, pT7:Maximin H5:His6 culture was at 0.45 OD. It was induced with IPTG and 0 hour sample was taken. At 5:00 pm, 3 hour sample was taken. Culture was let to grow overnight now.
7. Bacillus subtilis backbone received from Leiden university was inoculated in 20 mL culture.
8. Ligations on His-SUMO-Peptides with TetR cut backbones was done.
9. Ligations on His-SUMO-Peptides with pSB1C3(cut E+S) backbones was done.
10. Ligations on pBAD and AgrA:AgrC:pP2 was done.