Haynes Lab:Notebook/TB Biosensor/2012/09/13

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(Making crt Fusion Proteins by Gibson Assembly)
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(Making crt Fusion Proteins by Gibson Assembly)
 
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==Making crt Fusion Proteins by Gibson Assembly==
==Making crt Fusion Proteins by Gibson Assembly==
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* Ernesto PCR'd the overhangs onto the crt genes for Gibson assembly.  
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* Ernesto mixed the primers and templates below (to later PCR the overhangs onto the crt genes for Gibson assembly.)
[https://docs.google.com/spreadsheet/ccc?key=0Avv2LLPPodHAdDhDR1hFUXlvSFZleGd2RWhrcW03QXc  Primer details] with sequences
[https://docs.google.com/spreadsheet/ccc?key=0Avv2LLPPodHAdDhDR1hFUXlvSFZleGd2RWhrcW03QXc  Primer details] with sequences

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Making crt Fusion Proteins by Gibson Assembly

  • Ernesto mixed the primers and templates below (to later PCR the overhangs onto the crt genes for Gibson assembly.)

Primer details with sequences

Final Construct Template Forward Primer Reverse Primer
crtE - PLFlex - X crtE P001 P010
X- PLFlex - crtE crtE P015 P002
crtI - PLFlex - X crtI P005 P012
X- PLFlex - crtI crtI P017 P006
crtB - PLFlex - X crtB P007 P014
X- PLFlex - crtB crtB P019 P008
crtE - PLFlex - crtB PLFlex P009 P020
crtE - PLFlex - crtI PLFlex P009 P018
crtI - PLFlex - crtE PLFlex P011 P016
crtI - PLFlex - crtB PLFlex P011 P020
crtB - PLFlex - crtE PLFlex P013 P016
crtB - PLFlex - crtI PLFlex P013 P018


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