Team:HokkaidoU Japan/Notebook/aggregation Week 6

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">
 +
==August 6th==
 +
<div>
 +
==Ethanol precipitation==
 +
<p>
 +
Ethanol precipitation for digestion and gel extraction product.
 +
#Added 5 ul of NaoAc, 1.5 ul of glycogen and 125ul of 100% ethanol.
 +
#Centrifuged in 15000 rpm, 10 min at 4C.
 +
#Remove supernatant and added 220ul of 70% ethanol.
 +
#Centrifuged in 15000 rpm, 10 min at 4C.
 +
#Remove supernatant and air drying in room temperature then added 10 ul of DW.
 +
</p>
 +
 +
==Ligation==
 +
<p>
 +
We ligated pT7-RBS on pSB1K3 (digested Spe1) as vector and Ag43-dT (digested Spe1 and Xba1 and HindIII) as insert.
 +
 +
 +
{|class="hokkaidou-table-ligation"
 +
|-
 +
|pT7-RBS
 +
|1 ul
 +
|-
 +
|Ag43-dT
 +
|2 ul
 +
|-
 +
|DW
 +
|2 ul
 +
|-
 +
|Ligation Mighty Mix(TAKARA)
 +
|5 ul
 +
|-
 +
|Total
 +
|10 ul
 +
|}
 +
 +
 +
Ligation reaction time was written below.
 +
 +
{|class="hokkaidou-table-ligation"
 +
|-
 +
|Degree
 +
|Minute
 +
|-
 +
|16
 +
|30
 +
|-
 +
|65
 +
|10
 +
|-
 +
|4
 +
|Hold
 +
|}
 +
</p>
 +
 +
</div><div>
<!-- DO NOT EDIT UNDER THIS LINE @iTakeshi -->
<!-- DO NOT EDIT UNDER THIS LINE @iTakeshi -->

Revision as of 06:18, 6 August 2012



August 6th

Ethanol precipitation

Ethanol precipitation for digestion and gel extraction product.

  1. Added 5 ul of NaoAc, 1.5 ul of glycogen and 125ul of 100% ethanol.
  2. Centrifuged in 15000 rpm, 10 min at 4C.
  3. Remove supernatant and added 220ul of 70% ethanol.
  4. Centrifuged in 15000 rpm, 10 min at 4C.
  5. Remove supernatant and air drying in room temperature then added 10 ul of DW.

Ligation

We ligated pT7-RBS on pSB1K3 (digested Spe1) as vector and Ag43-dT (digested Spe1 and Xba1 and HindIII) as insert.

pT7-RBS 1 ul
Ag43-dT 2 ul
DW 2 ul
Ligation Mighty Mix(TAKARA) 5 ul
Total 10 ul


Ligation reaction time was written below.

Degree Minute
16 30
65 10
4 Hold


Retrieved from "http://2012.igem.org/Team:HokkaidoU_Japan/Notebook/aggregation_Week_6"