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CAROL
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WIKI CODING HERE
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CHINMOYEE
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EMILY
Going Back to Biobricking LuxOD47E
- Restriction digest products were mixed with 10x Orange Dye and visualized on a 1% agarose gel with Generuler 1.0 KB+ DNA Ladder and uncut plasmid of the same colonies as a negative conttrol. Gel photo is below.
- Lanes 1,3,5 and 7 are the restriction digest products for colonies 1-4 and lanes 2,4,6 and 8 are uncut plasmid for colonies 1-4.
- Analysis: There seems to have been no cutting with NotI as the band sin the digested lanes are the same as the bands in the uncut lanes. There are also some strange bands at about 12 KB which are much too big to be LuxOD47E. It looks like my gene may be lacking the NotI restriction sites, which indiactes that the BBK Ampliification Gradient PCR to Biobrick my gene if interest may not have been successful. We will proceed by returning to this PCR nad repeating it.
- Repeated PCR, see protocol on June 6th 2009.
- PCR Products were visualized on a 1% agarose gel run at 120 V with 1.0 KB GeneRuler DNA Ladder. See gel photo below.
- Lanes 1-12 contain LuxOD47E in pCR2.1-TOPO vector. Lane 13 is a negative control with only Mastermix + ddH2O.
- From this gel, it looks like our gene if interest, LuxOD47E is there as we see bads around 1.4 KB, as expected (gene is 1362 base pairs). We will proceed by isoltaing plasmid from the PCR product and performing a NotI Verification digest.
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FAHD
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IMAN
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JAMIE
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JEREMY
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KATIE
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KEVIN
Transformation
In order to verify the competency of KT1144 cells, I have transformed pBluescript into them. Competent KT1144 cells are needed to test the mutant circuits.
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MANDY
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PATRICK
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PRIMA
Bacterial Transformation
I shadowed Jeremy today. He was transforming the BBK ligated vector to competent cells (Top 10). I'm not sure exactly what he was transforming but I just hung out to learn the procedures and purposes of transformation.
I followed up with a few companies. BioAlberta has generously donated $1000.00 to the team. I wrote a Thank-You letter on behalf of the team and sent it to the President of BioAlberta.
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STEFAN
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VICKI
A sexy circuit discussion
Fahd and I worked together to propose a possible response circuit. We investigated the possiblity of using parts from the registry that can form a bioplastic, which could be useful for encapsulating cells that need to be inserted into the human body to - say - degrade a biofilm, but need to resist an immune and inflammatory response. Some considerations for our next discussion on this front will be whether or not this part works, and if so, what form the bioplastic comes in.
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