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Team:HokkaidoU Japan/Projects/asB0034/Method
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<div class="fig fig400 para"> | <div class="fig fig400 para"> | ||
<img src="https://static.igem.org/mediawiki/2014/0/02/HokkaidoU_antisenseB0034_overview07.png"> | <img src="https://static.igem.org/mediawiki/2014/0/02/HokkaidoU_antisenseB0034_overview07.png"> | ||
| - | <div>How to make antisense B0034 by primer annealing</div> | + | <div>Fig1. How to make antisense B0034 by primer annealing</div> |
</div> | </div> | ||
<div class="fig fig400 para"><p> | <div class="fig fig400 para"><p> | ||
<img src="https://static.igem.org/mediawiki/2014/6/63/HokkaidoU_antisenseB0034_overview08.png"> | <img src="https://static.igem.org/mediawiki/2014/6/63/HokkaidoU_antisenseB0034_overview08.png"> | ||
| - | <div>Using restriction enzyme, XhoI, NcoI, we made stem-antisense conplex. </div> | + | <div>Fig2. Using restriction enzyme, XhoI, NcoI, we made stem-antisense conplex. </div> |
</div> | </div> | ||
<h1><p>How to assay</p> | <h1><p>How to assay</p> | ||
<h4><p>we selected RFP and GFP for target genes. We used fluorophotometer to measure how antisense worked. The colonies transformed antisense RNA and target gene used to do assay.</p> | <h4><p>we selected RFP and GFP for target genes. We used fluorophotometer to measure how antisense worked. The colonies transformed antisense RNA and target gene used to do assay.</p> | ||
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<p>(1)To cultivate the colony in 4 mL LB culture for about 20 hours</p> | <p>(1)To cultivate the colony in 4 mL LB culture for about 20 hours</p> | ||
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<p>(4)To measure fluorescence after 9 hour </p> | <p>(4)To measure fluorescence after 9 hour </p> | ||
| - | + | <div class="fig fig800"> | |
| + | <img src="https://static.igem.org/mediawiki/2014/2/25/HokkaidoU_antisenseB0034_overview09.png"> | ||
| + | <div>Antisense B0034 is induce by IPTG</div> | ||
| + | </div> | ||
<!--end contents--> | <!--end contents--> | ||
Revision as of 07:58, 14 October 2014
How to synthesize antisense RNA
Antisense-RBS fragments were synthesized based on BioBrick by primer annealing. The sides of antisense fragment have scar and restriction enzymes XhoI, NcoI.
Fig1. How to make antisense B0034 by primer annealing
Fig2. Using restriction enzyme, XhoI, NcoI, we made stem-antisense conplex.
How to assay
we selected RFP and GFP for target genes. We used fluorophotometer to measure how antisense worked. The colonies transformed antisense RNA and target gene used to do assay.
(1)To cultivate the colony in 4 mL LB culture for about 20 hours
(2)To control turbidity up to 0.1 at OD600
(3)To cultivate the colony in 2 mL M9ZB culture for 9 hours (IPTG induces antisense RNA, addition 20 uL )
(4)To measure fluorescence after 9 hour
Antisense B0034 is induce by IPTG
