Literature study

Prof. Marjan De Mey and Charlotte Verniers assisted us during brainstorming and advised us during our literature study and the writing and presentation of our paper. Frederik De Bruyn also threw in some ideas during brainstorming.

The paper was written by the students of the UGent iGEM team. Presentations were also made by the students.

In the lab

Pieter Coussement, Frederik De Bruyn and Gert Peters gave us an introduction to common lab techniques. Frederik also gave us a tutorial on Clone Manager. We received safety training from Gilles Velghe who also made us familiar with the lab. Isabelle Maryns showed us how to safely handle ethidiumbromide. Safety forms were filled in by the students and Prof. De Mey.

Inbio (the UGent centre of expertise for industrial biotechnology and biocatalysis) let us make use of their lab and made sure all the necessary lab equipment was at our disposal. The whole Inbio staff was always ready to assist and advise us when we asked for help.

Inbio provided us with the strain bearing plasmids that contain the ccdA-gfp construct we used (E. coli DH5a + pTGD-ccdA-Pmb1-BCD7-GFPCmFRT). They provided the bacterial strain in which the knock-in was performed (E. coli MG1655 ΔendA DE3), strains containing plasmids with the T7ccdB fragment (E. coli DH5a + pXSpFRT-T7ccdB) and the strain we used for making P1 phage lysate (E. coli BW26547), along with a small dose of P1 lysate. Inbio provided us with a linear backup of the ccdA-gfp construct. They also constructed backup strains (E. coli MG1655 ΔendA DE3 ccdA-Pmb1GFP-CmFRT & E. coli MG1655 ΔendA DE3 ccdA-Pmb1GFP) in which CIChE could be performed if other experiments failed.

Primers starting with MDM and CGL were designed by Prof. Marjan De Mey. Primers beginning with oMEMO were designed by Inbio. Primers starting with iGEM were designed by students of the UGent iGEM team. Pieter Coussement also designed some primers for a reverse mutation and provided us with the protocol for this back mutation.

All the labwork was conducted by the students of the UGent team. Labwork over the two months was distributed between all students. All results were analysed by the students, especially Jules and Tine. We did however receive invaluable insights from our instructors: Prof. Marjan De Mey, Pieter Coussement, Frederik De Bruyn and Gert Peters.

Other

The following was also accomplished by the students:

Tine and Marlies constructed the wiki page. All texts on the wiki were written by students. Tim made the little movie explaining our project. Eva created our logo. Posters, flyers & surveys were made by the students. Survey results were analysed by Eline. All figures without reference were created by students.

Concerning ethics:

Renate and Tine interviewed the following professional scientists who helped us gain more insight in several ethical issues: Prof. Filip Buekens, Prof. Geert De Jaeger, Prof. Tom Desmet, Prof. Wim Soetaert, Prof. Els Van Damme. Texts were written by Renate and Eva. Frederik De Bruyn brought us in contact with Prof. Buekens and joined the discussion.