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Team:UEA-JIC Norwich/Weekfour
From 2011.igem.org
Monday 4th july
In the morning we ran a gel of our PCR products from Friday to test whether we had managed to amplify the specific gene we wanted. We also spent more time planning the UK iGEM get together, and decided on the protocol we would follow when transforming moss.
Tuesday 5th july
The gel from yesterday showed no DNA products from our PCR. We performed another PCR procedure using an adapted protocol and a different polymerase (phusion polymerase, as this has a proof reading ability and is therefore better suited for cloning). However, a gel from this was also unsuccessful. We returned to the drawing board and found a potential fault in the reverse primer we'd designed. Having corrected the error, we plan to order the adapted primer tomorrow. In light of specific numbers for the UK conference we began booking rooms and caterers. Having decided on a selection marker for algae transformations (arginine biosynthesis) we secured some arginine and autoclaved it ready for use tomorrow.
Wednesday 6th July 2011
It was Gurdeep’s birthday but no one cared. Algae plating was performed under extremely sterile conditions as the possibility of contamination was high. Using a sterile stick we inoculated some of our sent Chlamydomonas sample and then placed it on a TAP plate, left to grow for at least seven days.
Thursday 7th July 2011
The appointed members of the algae group get into gear by growing some algae cultures, this was done under strict sterile conditions and then left in a continuously shaking in the incubator at 25 degrees Celsius at 200rpm for roughly a week.
Friday 8th July 2011
Ben Jevans performed another PCR on the miniprep products he had done during the week. The outcome of the PCR was ran on gel with dire success.
