Team:EPF Lausanne/Calendar/24 October 2013

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Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface Display
CLoning INP_EYFP_Streptavidin (IYS) and INP_Streptavidin_EYFP (ISY)

1) First PCRs (10µl)
- PCR to re-amplify the streptavidin from BBa_K283010 miniprep
- PCR to amplify a streptavidin sequence from BBa_K523010 miniprep but without the stop codon at the end
- PCR to linearize BBa_K523013 and add linker for INP_EYP_Streptavidin construct
- PCR to linearize BBa_K523013 and add linker for INP_Streptavidin_EYFP construct
- 0.8% Gel electrophoresis to verify the PCRs => The results were not so good for the streptavidin; we add amplification in our controls but not is the expected PCR. For the linear backbone we forgot to put the appropiate controls.

2) Second PCRs (50 µl)
- PCR (1) to re-amplify the streptavidin form BBa_K283013 intial PCR (we did a PCR from a PCR because we thought the miniprep was not good anymore)
- PCR (2) to amplify streptavidin ssequence from BBa_K523010 intial PCR thus removing the stop codon at the end
- PCR (3) to linearize BBa_K523013 and add linker for INP_EYP_Streptavidin construct
- PCR (4) to linearize BBa_K523013 and add linker for INP_Streptavidin_EYFP construct
- 0.8% Gel electrophoresis to verify the PCRs with appropriate controls => the backbones PCR products are higher than the sizes expected but we decided to go on with it.
PCRs purification

Nanoparticles

- Prepare buffers for pro-MMP2 activation