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Team:HokkaidoU Japan/RBS/Methods
From 2013.igem.org
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<img src="https://static.igem.org/mediawiki/2013/a/ab/HokkaidoU_RBS_methods1_800.png"> | <img src="https://static.igem.org/mediawiki/2013/a/ab/HokkaidoU_RBS_methods1_800.png"> | ||
<div>fig.1: oligos; RED: enhancer sequence, BLUE: SD sequence.</div> | <div>fig.1: oligos; RED: enhancer sequence, BLUE: SD sequence.</div> | ||
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<img src="https://static.igem.org/mediawiki/2013/9/9e/HokkaidoU_RBS_methods2_400.png"> | <img src="https://static.igem.org/mediawiki/2013/9/9e/HokkaidoU_RBS_methods2_400.png"> | ||
<div>fig.2: RBS construction</div> | <div>fig.2: RBS construction</div> | ||
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<img src="https://static.igem.org/mediawiki/2013/2/28/HokkaidoU_RBS_methods3_400.png"> | <img src="https://static.igem.org/mediawiki/2013/2/28/HokkaidoU_RBS_methods3_400.png"> | ||
<div>fig.3: our parts</div> | <div>fig.3: our parts</div> | ||
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Revision as of 00:36, 28 September 2013
Maestro E.coli
RBS
RBS family parts
We constructed new RBS family, SD2, SD4, SD6, SD8. These RBSs have Enhancer sequence (GCTCTTTAACAATTTATCA) and SD sequence (SD2:GG, SD4:GAGG, SD6:AGGAGG, SD8:TAAGGAGG). We constructed SD8 from synthetic oligos (forward:SD8-f, reverse:SD8-r). We constructed SD2, SD4, SD6 by PCR (forward:EX-f, reverse:SD2-r, SD4-r, SD6-r, template:SD8).
fig.1: oligos; RED: enhancer sequence, BLUE: SD sequence.
fig.2: RBS construction
fig.3: our parts
Assay
We ligated TetR repressible promoter (pTet), each of the new RBSs', LacZα and double terminator. Using this construct we performed β-Galactosidase assay.
