Team:HokkaidoU Japan/Notebook/aggregation Week 11
From 2012.igem.org
(Difference between revisions)
(10th digestion) |
(10th digestion recipe) |
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{|class="hokkaidou-table-digestion" | {|class="hokkaidou-table-digestion" | ||
|- | |- | ||
- | |DNA solution ( | + | |DNA solution ( 35ng/ul) |
- | | | + | |17 ul |
|- | |- | ||
- | | | + | |EcoRI |
|1 ul | |1 ul | ||
|- | |- | ||
- | | | + | |SpeI |
|1 ul | |1 ul | ||
|- | |- | ||
- | | | + | |10xH buffer |
|3 ul | |3 ul | ||
|- | |- | ||
|DW | |DW | ||
- | | | + | |8 ul |
+ | |- | ||
+ | |Total | ||
+ | |30 ul | ||
+ | |} | ||
+ | |||
+ | |||
+ | |||
+ | Insert2 (Ag43-dT-pSB1AK3) | ||
+ | {|class="hokkaidou-table-digestion" | ||
+ | |- | ||
+ | |DNA solution ( 35ng/ul) | ||
+ | |25 ul | ||
+ | |- | ||
+ | |XbaI | ||
+ | |1 ul | ||
+ | |- | ||
+ | |NotI | ||
+ | |1 ul | ||
+ | |- | ||
+ | |10xK buffer | ||
+ | |1.5 ul | ||
+ | |- | ||
+ | |100xBSA | ||
+ | |0.3ul | ||
+ | |- | ||
+ | |DW | ||
+ | |1.5 ul | ||
|- | |- | ||
|Total | |Total | ||
Line 35: | Line 62: | ||
- | Vector( | + | Vector(pSTV28) |
{|class="hokkaidou-table-digestion" | {|class="hokkaidou-table-digestion" | ||
|- | |- | ||
- | |DNA solution ( | + | |DNA solution ( 15ng/ul) |
- | | | + | |9 ul |
|- | |- | ||
- | | | + | |EcoRI |
|1 ul | |1 ul | ||
|- | |- | ||
- | | | + | |NotI |
|1 ul | |1 ul | ||
|- | |- | ||
|10xH buffer | |10xH buffer | ||
|2 ul | |2 ul | ||
+ | |- | ||
+ | |10xBSA | ||
+ | |0.2ul | ||
|- | |- | ||
|DW | |DW | ||
- | | | + | |7 ul |
|- | |- | ||
|Total | |Total | ||
Line 58: | Line 88: | ||
- | control ( | + | control (pT7-RBS-pSB1C3) |
{|class="hokkaidou-table-digestion" | {|class="hokkaidou-table-digestion" | ||
|- | |- | ||
- | |DNA solution (30 ng/ul) | + | |DNA solution (30~40 ng/ul) |
- | | | + | |10 ul |
|- | |- | ||
|XbaI | |XbaI | ||
Line 74: | Line 104: | ||
|- | |- | ||
|DW | |DW | ||
- | | | + | |6 ul |
|- | |- | ||
|Total | |Total | ||
Line 93: | Line 123: | ||
|- | |- | ||
|2 | |2 | ||
- | | | + | |70 |
- | | | + | |20 |
|- | |- | ||
|3 | |3 | ||
Line 100: | Line 130: | ||
|HOLD | |HOLD | ||
|} | |} | ||
+ | </p> | ||
</div></div> | </div></div> | ||
<!-- DO NOT EDIT UNDER THIS LINE @iTakeshi --> | <!-- DO NOT EDIT UNDER THIS LINE @iTakeshi --> |
Revision as of 05:08, 10 September 2012
September 10th
Digestion of eCFP-RBS-pSB1A2 and pBAD-RBS-pSB1A2
To make a construct of pBAD-RBS-eCFP-RBS-Ag43-dT-pSTV28 by 3piece ligation, we digested pBAD-RBS-eCFP-RBS-pSB1A2 with EcoRI & SpeI, Ag43-dT-pSB1AK3 with XbaI & NotI and pSTV28 with EcoRI and NotI. Then we digested pT7-RBS-pSB1C3 with XbaI & SpeI as a control for confirmation of the ability of restriction enzyme. Insert1 (pBAD-RBS-eCFP-RBS-pSB1A2)
DNA solution ( 35ng/ul) | 17 ul |
EcoRI | 1 ul |
SpeI | 1 ul |
10xH buffer | 3 ul |
DW | 8 ul |
Total | 30 ul |
Insert2 (Ag43-dT-pSB1AK3)
DNA solution ( 35ng/ul) | 25 ul |
XbaI | 1 ul |
NotI | 1 ul |
10xK buffer | 1.5 ul |
100xBSA | 0.3ul |
DW | 1.5 ul |
Total | 30 ul |
Vector(pSTV28)
DNA solution ( 15ng/ul) | 9 ul |
EcoRI | 1 ul |
NotI | 1 ul |
10xH buffer | 2 ul |
10xBSA | 0.2ul |
DW | 7 ul |
Total | 20 ul |
control (pT7-RBS-pSB1C3)
DNA solution (30~40 ng/ul) | 10 ul |
XbaI | 1 ul |
SpeI | 1 ul |
10xM buffer | 2 ul |
DW | 6 ul |
Total | 20 ul |
Number | Degree | Minute |
1 | 37 | 120 |
2 | 70 | 20 |
3 | 4 | HOLD |