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Team:BCCS-Bristol/Notebook
From 2009.igem.org
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===Week 1=== | ===Week 1=== | ||
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| + | ====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. | *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 | *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. | *Decided to use 3 reporter genes, 1 RBS, 1 High Copy plasmid backbone for now. | ||
| - | ====Reporters==== | + | |
| + | =====Reporters===== | ||
*RFP(Bba_E1010) | *RFP(Bba_E1010) | ||
*GFP(Bba_E1040) | *GFP(Bba_E1040) | ||
*LacZ(Bba_I732005) | *LacZ(Bba_I732005) | ||
| - | ====RBS==== | + | =====RBS===== |
*Bba_J61100 - From Anderson Family | *Bba_J61100 - From Anderson Family | ||
| - | ====Plasmid Backbone==== | + | =====Plasmid Backbone===== |
*BBa_J04450 ; pSB1A3 | *BBa_J04450 ; pSB1A3 | ||
Revision as of 09:20, 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
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.
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.
