Team:BCCS-Bristol/Notebook

From 2009.igem.org

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===Week 2===
===Week 2===
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====PCR====
*Primers finally arrived. Did PCR to amplify carrier genes.
*Primers finally arrived. Did PCR to amplify carrier genes.
*PCR worked. Restricted DNA and inserted into biobrick backbone pSB1A3.
*PCR worked. Restricted DNA and inserted into biobrick backbone pSB1A3.
 +
 +
====In frame protein fusions====
*Started working on finding an easy assembly method for in-line 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.
*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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Revision as of 09:18, 24 July 2009

BCCS-Bristol
iGEM 2009


Contents

Week Zero

  • Familiarising with standard lab procedures for the past week (bacterial culture growth, restriction enzyme usage, agarose gel electrophoresis).
  • Will start designing some biobricks for the project today.

Week 1

  • 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
  • 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.

Week 2

PCR

  • Primers finally arrived. Did PCR to amplify carrier genes.
  • PCR worked. Restricted DNA and inserted into biobrick backbone pSB1A3.

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.