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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. | ||
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| + | ====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
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.
