"
Team:HokkaidoU Japan/Projects/asB0034/Method
From 2014.igem.org
(Difference between revisions)
Kakurehana (Talk | contribs) |
|||
| Line 94: | Line 94: | ||
<!--begin contents--> | <!--begin contents--> | ||
| + | <h1>How to synthesize antisense RNA | ||
| + | <h4><p>Antisense-RBS fragments were synthesized based on BioBrick by primer annealing. The sides of antisense fragment have scar and restriction enzymes XhoI, NcoI.</p> | ||
| + | |||
| + | <h1>How to assay | ||
| + | <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> | ||
| + | |||
| + | <p>(1)To cultivate the colony in 2 mL LB culture for 9 hours</p> | ||
| + | <p>(2)To control turbidity up to 0.1 at OD600</p> | ||
| + | <p>(3)To cultivate the colony in 2 mL M9ZB culture for 9 hours (IPTG induces antisense RNA, addition 20 uL )</p> | ||
| + | <p>(4)To measure fluorescence after 9hours, 11hours, 13hours </p> | ||
Revision as of 03:35, 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.
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 2 mL LB culture for 9 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 9hours, 11hours, 13hours
