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Team:HokkaidoU Japan/Notebook/August24
From 2010.igem.org
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{{Template:HokkaidoU_Japan}}<div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/August23|August 23]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/August25|August 25]]</div></div> | {{Template:HokkaidoU_Japan}}<div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/August23|August 23]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/August25|August 25]]</div></div> | ||
| - | = | + | =Check to see if digestion visualization Primers Work= |
| - | [[Image:HokkaidoU Japan 20100824a.jpg|200px|right|thumb| | + | |
| - | * | + | [[Image:HokkaidoU Japan 20100824a.jpg|200px|right|thumb|Electrophoresis after restriction enzyme digestion]] |
| + | |||
| + | * All 4 of PCR products were purified via Microcon YM-10 | ||
| + | |||
{| class="protocol" | {| class="protocol" | ||
|'''Lane''' | |'''Lane''' | ||
| Line 23: | Line 26: | ||
|Promoter | |Promoter | ||
|} | |} | ||
| - | + | Promoter band in lane 6 wasn't visible | |
| - | + | * This vector didn't have a primer annealing site for our new primers | |
| - | * | + | |
=Ligation Mixtureのチェック= | =Ligation Mixtureのチェック= | ||
Revision as of 05:46, 22 September 2010
since 13 Jul 2010
since 1 Aug 2010
Check to see if digestion visualization Primers Work
- All 4 of PCR products were purified via Microcon YM-10
| Lane | DNA |
| 1 | TSUDA Marker I |
| 3 | RBS |
| 4 | GFP |
| 5 | double terminator |
| 6 | Promoter |
Promoter band in lane 6 wasn't visible
- This vector didn't have a primer annealing site for our new primers
Ligation Mixtureのチェック
Ligation Mixture「I」とLigation Mixture「M」について
- PCRで増やしてゲル抽したpSB1C3 2 uLにLigation Mixture 4 uLをいれ，それぞれのTransformationした
- LB 200 uLを加え，100 uLずつをChloramphenicol 34 ug/uLと 12 ug/uLの培地にまいた
いろいろ確認
| Lane | DNA |
| 2 | λ/Hind III |
| 3 | pSB1C3どうしのLigation |
| 4 | ラオリプライマーによる1-2NのPCRのろ過産物（上） |
| 5 | ラオリプライマーによる1-2NのPCRのろ過（下） |
| 6 | λ/Hind III |
| 7 | ゲル抽したpSB1C3 |
レーン３でバンドが見られなかった．
- ダイマー，テトラマーなど，さまざまな断片ができ，薄くなってしまった？
レーン７は薄いがバンドが確認できた
- ゲル抽に問題はない
