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.
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.
Every member of the team signed up to be a STEM ambassador (please see outreach page). Angela Carpenter arrived at the JIC in the morning and gave an induction pack to each member of the team. She then took us through a slideshow presentation and gave us useful advice and information into working with children. She also took personal details from each member of the team in order to complete CRB checks.
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.
The appointed members of the algae group get into gear by growing some algae cultures, this was done under sict sterile conditions and then left in a continuously shaking in the incubator at 25 degrees Celsius at 200rpm for roughly a week.
Ben Jevans performed another PCR on Kimberley, on the miniprep products he had don during the week. The outcome of the PCR was ran on gel with dire success.