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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
Verifictaion Digest of J13002-LuxOD47E-B0015
- Today I did a minprep of my overnight cultures of J13002-LuxOD47E-B0015. I took the concentration and did a restriction digest with XbaI and PstI. I ran this on a gel this afternoon with uncut plasmids of each colony as a control as well as LuxOD47E BBK (uncut) and J13002-LuxOD47E (uncut) as size controls. See gel photo below.
Lane 1- J13002-LuxOD47E-B0015 Trial 2, C1 cut with XbaI and PstI
Lane 1- C 1, uncut
Lane 3- C2, Cut
Lane 4- C2, Uncut
Lane5- C3, Cut
Lane 6- C3, Uncut
Lane 7-LuxOD47E BBK (Uncut)
Lane 8- J13002-LuxOD47E (Uncut)
Outreach/ Ethics
- Stefan, Fahd and I met to talk about Ethics this morning to make a plan of what needs to be done for the Erhics conference. We're planning the conference for some time in late September/ early October, but we want to start inviting people now. So today we made a list of possible speakers as well as people that we will invite just to come and listen.
- We decided that the first step would be to familiarize ourselves with Second Life. So, this afternoon I made an account on SecondLife and explored a little. I walked through the boards with instructions on them to learn how to talk, move, ect. Mandy then gave me access to our Island, so I got a chance to walk around and look at some of the stuff that our team has been looking on. Tomorrow I'm going to explore an ethics area that Mandy knows about as well as try to find some conferences to sit in on to get a better idea of how they are run. Fahd and I need to get planning our Ethics conference so I think it's a really good idea to become familiar with what we can do in Second Life so that we can decide where we want to have it, what we need to build for it, how we're going to advertise it within Second Life, etc.
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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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