Team:UEA-JIC Norwich/Weekfour

From 2011.igem.org

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<br>Monday 4th july
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<br>Monday 27th June
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Today, we spent the morning researching moss and received our sample of algae in a nappy! A slideshow of photos was embedded onto the home page of the wiki and then we had lunch. After lunch we carried out transformations with 5 different biobricks, where the details can be viewed in the lab journal.
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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.
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<br>Tuesday 28th June 2011
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<br>Tuesday 5th july
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Today a transformation was carried out using the dual regulated light response system Biobrick, all hoping that it will be a potentially useful part in our future plans.
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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.
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<br>Wednesday 29th June 2011
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<br>Wednesday 6th July 2011
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The transformed E.coli was recuperated from its overnight incubation and then a single colony was picked off the plate and placed into LB-broth to grow overnight ready for tomorrow.
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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.
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<br>Thursday 30th June 2011
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<br>Thursday 7th July 2011
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Today the team attempt their first ever miniprep of the culture grown on the previous day, it may of been done slowly but more importantly carefully and precise. A glycerol stock was also made of the grown culture so we may preserve the the cells to be used another day.
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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.
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<br>Friday 1st July 2011
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<br>Friday 8th July 2011
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In the morning we ran a gel of our Miniprep of the BBA_M30109 transformed E.coli cells. The Gel showed that we had indeed successfully isolated the plasmid from our cells, and so in the evening (LATE into the evening) we ran PCR protocol to try and amplify the specific gene we wanted from the plasmid (Cph8) We also continued to arrange the UK meet up, adding up numbers that were coming and trying to find speakers. We also added some finishing touches to the logo and placed it onto a business card for the team.
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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.

Revision as of 17:14, 20 September 2011

University of East Anglia-JIC

UNIVERSITY OF EAST ANGLIA-JOHN INNES CENTRE


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.

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