IGEM:Groningen/Notebook/iGEM 2011/2011/09/07

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7-9-11



Project is almost comming to an end, also the amount of taq polymerase is getting very low. Searching for more taq:
Borrowed dreamtaq, in the end there was enough normal taq left, but Berke needed to use the dreamtaq.

Colony PCR of PBAD-RBS-cI-LVA (2×9),PBAD-RBS-cI-LVA-DT (1×9), PBAD-LasR-LVA-DT (1×9), PBAD-RBS-GFP-DT(2×9)
Mastermix:
Taq 10× buffer: 116μl
dNTPs 10mM: 23.2μl
MgCl2: 69.6μl
Forward Biobrick primer: 23.2μl
Reverse Biobrick primer: 23.2μl
Taq DNA polymerase: 5.8μl
MQ water: 899μl

PCR conditions:
Preheated lid: 111°C
Denaturation: 94°C for 10 min.
Cycle 33×:
denaturation: 94°C for 30s.
annealing: 60°C for 30s.
Extension: 72°C for 2,5 min.
Final extension: 72°C for 10 min.
Store infinite at 4°C

Analyse on a 1% TBE agarose gel.

After analysis of the 3 gels, make two new ones and put Berke's and Christophs samples in there.
Analyse on a 1% TBE agarose gel

During the day: working on the wiki and making phone call for the church choir's general recording of the song.
Also work on the presentations for the schools.
Start making a dayplan and also work out the practicum thing for the students (how much of the solutions do we need and what
do I need to buy and make sure there is a waterbath etc.

Clones of the transformants of yesterday checked today with colony PCR look alright!:)
These colonies were grown overnight in LB medium with Cm (except 1 PBAD-RBS-GFP-DT in ampicillin vector)
PBAD-RBS-cI-LVA:1, PBAD-RBS-cI-LVA(2):1,PBAD-RBS-cI-LVA-DT: 1+3, PBAD-LasR-LVA-DT 3+5 ,PBAD-RBS-GFP-DT: 6,PBAD-RBS-GFP-DT(2): 4

Gel with Berke's and Christoph's samples finished: made a photo on the gel doc and send it to them by e-mail

At 5 there will be a presentation meeting to let the presenters practice their presentation for tomorrow's GBB symposium!






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