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CAROL
Wiki Updates
- Updated the wiki pages today, so there is full descriptions for all the parts of our project. As well, started working on the protocols for our wiki today.
- The transformation with pCS26 vector and the sigma 70 promoters ligated by T4 Invitrogen Ligase showed small colonies on plates late in the afternoon, left overnight in the incubator.
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CHINMOYEE
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EMILY
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FAHD
Second Life Conference, July Newsletter, Education and Outreach:
Today I sent out July letters to the Following comapnies:
1)Nexen Inc.
2)New England Biolabs
3)Alberta Ingenuity Fund
4)Boheringer Ingleheim Canada
5)Apotex Canada
My Ethics teammates and I today discussed some ways to conduct the Ethics conference on synthetic biology in the online virtual world called Second Life.
I also had a meeting with our education and outreach team. We discussed ways to promote iGEM in high schools and universities.
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IMAN
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JAMIE
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JEREMY
Plasmid Isolation of cl lambda and Colony PCR of LuxPQ-B0015-R0040-LuxOU-B0015 in psB1AC3
Plasmid was isolated from an overnight culture of cl lamba using Sigma GenElute Mini Prep Kit. The concentration and purity of the isolated plasmid was then measured with the NanoDrop spectrophotometer. Restriction digest was performed with XbaI and PstI in React 2 (Invitrogen) buffer for 2 hours at 37ºC. Cut plasmid and uncut plasmid were then run on a 1.0% agarose gel, no bands were seen for either column. The isolated plasmid was then specked again, to verify the initial concentration, which turned out to be accurate. An overnight Restriction digest was then set up as described above; however, 600ng of DNA were used instead of 200ng.
A colony PCR was set up with colonies of the construction LuxPQ-B0015-R0040-LuxOU-B0015 in psB1AC3 as the template. Seven colonies were chosen. This PCR was performed with BBK CP F/R primers (these anneal on the BBK vector just outside the cloning site) and LuxPQ F / LuxOU R primers (these are gene specific primers). Due to the long extension time for this PCR (6min and 20 seconds) it is run overnight and then held at 4ºC upon completion. For PCR conditions, refer to PCR performed on July 27 2009. Restreaks of the seven colonies were made and overnight liquid cultures were made so that plasmid can be isolated tomorrow.
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KATIE
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KEVIN
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MANDY
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PATRICK
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PRIMA
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STEFAN
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VICKI
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