Team:BCCS-Bristol/Notebook/Week 1

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BCCS-Bristol
iGEM 2009



Contents

Week 1

Canditate Proteins for Biobricks

  • Isolated 3 canditate proteins to act as carriers for our biobricks. These are FhuA,Fiu & OsmE. Started to design primers to amplify the selected genes via PCR.
  • Primers designed and ordered. Waiting for their arrival to do PCR! :D

Reporters,RBS,Backbones

  • Decided to use 3 reporter genes, 1 RBS, 1 High Copy plasmid backbone for now.
Reporters
*RFP(Bba_E1010)
*GFP(Bba_E1040)
*LacZ(Bba_I732005)
RBS
*Bba_J61100 - From Anderson Family
Plasmid Backbone
*BBa_J04450 ; pSB1A3
  • Tried to extract DNA from the iGEM biobricks and transfrom into bacteria.
  • Transformations do not work properly with non-commercial E.coli strain (XL-1).
  • Transformations worked the 2nd time round with commercial Nova Blue E.coli Strain. DNA samples in toolkit must be of low concentrations!
  • Regrew bacterial colonies to amplify DNA of reporters,RBS,backbone.
  • Miniprepped the DNA of Reporters,RBS,plasmid backbone and made glycerol stocks.
  • Realised that we are faced with a problem when wanting to assemble biobricks for protein fusions.