Team:EPF Lausanne/Calendar/5 September 2013


Taxi.Coli: Smart Drug Delivery iGEM EPFL


Cell Surface Display
- Sending Gibson Alive Miniprep product for sequencing using VR and VF2 iGEM primers.
- Sending Gibson Dead and iGEM miniprep for sequencing using VF2 primer once we didn't do it last time.
New Strategy
Primer designing for a new strategy to create the following fusion proteins: INP_YFP_strepta and INP_strepta_YFP.


PCR of the two pH sensitive Promoters and the constitutive promoter from iGEM
-We used the same program as the day before in order to amplify the promoters. For the hya and the Cad promoters we put 3 min initial denaturation as they were contained in genomic DNA. Again the PCR was successful

Purification of PCR products
-After PCR we purified the products with the qiagen Kit according to the manufacturers instructions. As the pH promoter from iGEM is only 30 bp it was removed during purification. So we decided to amplify it again but with primers that had overhangs with the backbone, in order to have an insert of about 100 bp. The final concentrations of the purifications were all very low. I still did a DpnI digestion with 8 ul DNA and then a Gibson assembly but it did not work. As I had done only a 20 ul reaction of the PCR, I had no more Product left so I decided to do the PCR again but with a volume of 50 ul.


ELISA-like assay
Continuing with the ELISA-like assay.