Team:BCCS-Bristol/Notebook

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{{:Team:BCCS-Bristol/Header}}
 
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===Week Zero===
 
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*Familiarising with standard lab procedures for the past week (bacterial culture growth, restriction enzyme usage, agarose gel electrophoresis).
 
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*Will start designing some biobricks for the project today.
 
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===Week 1===
 
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====Canditate Proteins for Biobricks====
 
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*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.
 
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*Primers designed and ordered. Waiting for their arrival to do PCR! :D
 
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====Reporters,RBS,Backbones====
 
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*Decided to use 3 reporter genes, 1 RBS, 1 High Copy plasmid backbone for now.
 
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=====Reporters=====
 
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*RFP(Bba_E1010)
 
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*GFP(Bba_E1040)
 
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*LacZ(Bba_I732005)
 
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=====RBS=====
 
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*Bba_J61100 - From Anderson Family
 
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=====Plasmid Backbone=====
 
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*BBa_J04450 ; pSB1A3
 
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*Tried to extract DNA from the iGEM biobricks and transfrom into bacteria.
 
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*Transformations do not work properly with non-commercial E.coli strain (XL-1).
 
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*Transformations worked the 2nd time round with commercial Nova Blue E.coli Strain. DNA samples in toolkit must be of low concentrations!
 
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*Regrew bacterial colonies to amplify DNA of reporters,RBS,backbone.
 
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*Miniprepped the DNA of Reporters,RBS,plasmid backbone and made glycerol stocks.
 
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*Realised that we are faced with a problem when wanting to assemble biobricks for protein fusions.
 
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===Week 2===
 
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====PCR====
 
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*Primers finally arrived. Did PCR to amplify carrier genes.
 
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*PCR worked. PCR products (3 candidate proteins) and inserted into biobrick backbone pSB1A3.
 
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====In frame protein fusions====
 
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*Started working on finding an easy assembly method for in-line protein fusions.
 
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*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.
 

Latest revision as of 00:01, 21 October 2009