Team:BCCS-Bristol/Notebook/Week 2

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Revision as of 12:43, 25 September 2009

Team:BCCS-Bristol:Header

Team:BCCS-Bristol:NotebookHeader


Week 2

PCR

  • Primers finally arrived. Did PCR to amplify carrier genes.
  • PCR worked. PCR products (3 candidate proteins)ligated onto biobrick backbones pSB1A3 and pSB1A2 (contains RBS BBa_J61100).
  • 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!!)
  • Most transformations are successful. Used transformed colonies to prepare liquid cultures so that we can proceed with minipreping them.

In frame protein fusions

  • Started working on finding an easy assembly method for in-line protein fusions.
  • 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.
  • Prepared different versions of this Bioscaffold to be ordered and tested in the lab for actual functionality.