From 2009.igem.org
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| - | {{:Team:BCCS-Bristol/Header}}
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| - | {{:Team:BCCS-Bristol/NotebookHeader}}
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| - | ===Week 2===
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| - | ====PCR====
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| - | *Primers finally arrived. Did PCR to amplify carrier genes.
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| - | *PCR worked. PCR products (3 candidate proteins)ligated onto biobrick backbones pSB1A3 and pSB1A2 (contains RBS BBa_J61100).
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| - | *The plasmid backbones with the genes of interest inserted into them were used to transform the XL1-BLUE E.coli strain (although not competent enough compared to NovaBlue cells they are much cheaper!!)
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| - | *Most transformations are successful. Used transformed colonies to prepare liquid cultures so that we can proceed with minipreping them.
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| - | ====In frame protein fusions====
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| - | *Started working on finding an easy assembly method for in-line protein fusions.
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| - | *Developed the design for a Bioscaffold-Linker transformer family (inspired by Bioscaffolds). Should allow fusions of proteins and all RFC10 biobricks in-frame after using Bioscaffold specific restriction enzymes.
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| - | *Prepared different versions of this Bioscaffold to be ordered and tested in the lab for actual functionality.
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| - | [[Image:BCCS_Carrier_proteins_PCR.jpg|center|500px|frame|Novel protein additions were cloned from the MG1655 genome with PCRt. Lane 1 shows the presence of fhuA, lane 2 fiu and lane 3 osmE. The ladder is Lambda-DNA HindIII digest from NEB (In descending order; 23130, 9416, 6557, 4361,2322, 2027. Carrier proteins were then placed onto pSB1A3 plasmid backbones to create biobricks novel protein carriers.]]
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Latest revision as of 23:47, 20 October 2009
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