June

Week 1

Monday

Tuesday

Preparation of freezable competent cells (M15): 40 alicos of 100µL
Test of the competent cells.

Wednesday

Begining of ‘intra-cell ROS concentration’ modelisation.

Transfection of competent cells with the biobricks :
- pLac
- mCherry
- cl Inverter
- RBS (Elowitz 1999)
- GFP

First contact with Arduino : the electronic device that will be used as a controler, just as the Cambridge 2010 iGEM team.
Culture of the BBs k174000 and K592004/5/6

Thursday

Failure with the transfections.
Other problem with the petri dishes : everything grew, even the WT colonies that should have died because of antibiotics. Maybe the antibiotics are too old since they were used by Grenoble iGEM Team of 2012

Friday

Preparation of freezable competent cells (WT)(40 alicos of 100µL)
Same transfections than wednesday, with fresh antibiotics.

PCRs of pAraBad, mCherry and KillerRed, only pAraBAD worked.

Team meeting
TODO List :
- Find a name
- Begin construction of :
->pLac-RBS-YF1-fixJ
->pFixK2-RBS-GFP
- glycerol-stock BBs
- E-glometer, caracterisation of LED

Week 2

Little Summary with Keywords

Monday

PCRs of mCherry and KR (did not work). Is there a problem with the primers or with the DNA templates? Need to work on that
Put the transformed cells on petri dishes with the right antibiotics
Receive the TSL230RD photodiodes
Begin the test of Arduino (digital output) by turning on and off a LED and using a Graphical User Interface (GUI) to control the intensity

Construction of the pQE30::KillerRed vector

Incubated a 50mL overnight culture of E. coli M15[pRep4] cells (Qiagen, Venlo, Netherlands), containing the pQE30::αSNAP vector, in preparation for midiprep.

Tuesday

Midi-prepped pQE30::αSNAP. The DNA sample concentration was only of 10 ng/µL. The experiment has to be re performed.

Incubated a 10mL overnight culture of E. coli M15[pRep4-pQE30::αSNAP] cells, in preparation for miniprep.

Wednesday

Construction of the pQE30::KillerRed vector
Mini-prepped pQE30::αSNAP/pRep4 and got a 98,9 ng/µL DNA sample.

Digested pQE30::αSNAP with BamHI and KpnI restriction enzymes. Separation of the gene of non interest (αSNAP) from the pQE30 vector backbone by gel electrophoresis (1.2 % agarose, 30 min, 135V).

Thursday

Friday

Week 3

Week 4