Fatty acids

Fish normaly provides a great amount of essential fatty acids. We wanted to clone ∆9, ∆12 & ∆15 fatty acid desaturases into separate plasmids by the use of USER cloning. We wanted to use USER cloning to save some time and to ease the cloning process. Ahead of this cloning, we needed the desaturases separately.

We tried running several USER PCRs on ∆9 desaturase iGEM part but all were unsuccessful.

We received the ∆12 desaturase (FAT-2) which originated from Caenorhabditis elegans. We ran a PCR which was successful. Thus we cloned it into a PSB1C3 plasmid and it can now be found in parts registry under BBa_K1475002. Afterwards we tried running USER PCR on FAT-2 which was unsuccesful.

We tried to run a colony PCR on the bacterium Synechocystis sp. PCC6803, with Phusion polymerase, in order for us to get the ∆15 desaturase. Unfortunately, this was unsuccessful.

After several failed attempts to run USER PCRs, we realized that we needed to prioritize and continue without cloning ∆9, ∆12 & ∆15 into the separate plasmids, with the use of USER cloning.