Team:HokkaidoU Japan/RBS/Methods

From 2013.igem.org

(Difference between revisions)
Line 24: Line 24:
<div class="fig800">
<div class="fig800">
   <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>
</div>
</div>
<div class="fig400 para">
<div class="fig400 para">
   <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>
</div>
</div>
Line 35: Line 35:
<div class="fig400 para">
<div class="fig400 para">
   <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>
</div>
</div>
<div class="clearfix"></div>
<div class="clearfix"></div>

Revision as of 23:45, 27 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.