Team:HokkaidoU Japan/Notebook/aggregation Week 5

From 2012.igem.org

(Difference between revisions)
(Created page with "{{Team:HokkaidoU_Japan/header}} {{Team:HokkaidoU_Japan/nav.notebook}} <div id="hokkaidou-column-main"> <!-- DO NOT EDIT OVER THIS LINE @iTakeshi --> <!-- DO NOT EDIT UNDER THIS ...")
Line 3: Line 3:
<div id="hokkaidou-column-main">
<div id="hokkaidou-column-main">
<!-- DO NOT EDIT OVER THIS LINE @iTakeshi -->
<!-- DO NOT EDIT OVER THIS LINE @iTakeshi -->
 +
<div class="hokkaidou-notebook-daily">
 +
==July 30th==
 +
<div>
 +
==digestion==
 +
<p>
 +
I tried to digeste the mini-prep products again, and after Ethanol precipitation, we use electrophoresis.
 +
[[image:HokkaidoU2012_120730_ag43-f1-e-s.jpg|thumb|digestion result]]
 +
 +
</p>
 +
==transformation==
 +
<p>
 +
Now, I think like this.</ br>
 +
The E. coli which we use transformation is BL21(DE3)pLysS.
 +
So, the E. coli could multiplicationincrease on LBC plate.</ br>
 +
I'm going to do transformation , use DH5α.
 +
 +
Transformation of plasmid DNA ligated in 25th (pT7-RBS-Ag43-dT on pSB1K3) in E. coli(DH5α).
 +
#Added 2ul of plasmid DNA to 50ul of thawed competent cells on ice.
 +
#Incubated on ice for 30min.
 +
#Added 200ul of LB then incubated the cells for 2 hours at 37C.
 +
#Prepared and Labeled two petri dishes with LBK.
 +
#Plate 200ul of the transformation onto first dish and spread.
 +
#Added 450ul of LB to 50ul of the transformation and plated 200ul of it onto second dish and spread.
 +
#Incubated the plates at 37C for over 30hrs.
 +
 +
 +
 +
<!-- DO NOT EDIT UNDER THIS LINE @iTakeshi -->
<!-- DO NOT EDIT UNDER THIS LINE @iTakeshi -->

Revision as of 12:24, 30 July 2012

July 30th

digestion

I tried to digeste the mini-prep products again, and after Ethanol precipitation, we use electrophoresis.

digestion result

transformation

Now, I think like this.</ br> The E. coli which we use transformation is BL21(DE3)pLysS. So, the E. coli could multiplicationincrease on LBC plate.</ br> I'm going to do transformation , use DH5α. Transformation of plasmid DNA ligated in 25th (pT7-RBS-Ag43-dT on pSB1K3) in E. coli(DH5α).

  1. Added 2ul of plasmid DNA to 50ul of thawed competent cells on ice.
  2. Incubated on ice for 30min.
  3. Added 200ul of LB then incubated the cells for 2 hours at 37C.
  4. Prepared and Labeled two petri dishes with LBK.
  5. Plate 200ul of the transformation onto first dish and spread.
  6. Added 450ul of LB to 50ul of the transformation and plated 200ul of it onto second dish and spread.
  7. Incubated the plates at 37C for over 30hrs.
</div>
</div> </div> </div>

Retrieved from "http://2012.igem.org/Team:HokkaidoU_Japan/Notebook/aggregation_Week_5"