Team:HokkaidoU Japan/Notebook/August19
From 2010.igem.org
(Difference between revisions)
(→Ligation) |
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!パーツ | !パーツ | ||
- | ! | + | !By comparison to marker |
!size (bp) | !size (bp) | ||
- | ! | + | !(ng) required |
- | ! | + | !(uL) used |
|- | |- | ||
|Vector | |Vector | ||
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|style="border-top:1px solid #996;"|'''3.3 uL''' | |style="border-top:1px solid #996;"|'''3.3 uL''' | ||
|} | |} | ||
- | + | Ligation and transformation follows same protocol as previously | |
=competencyを求める= | =competencyを求める= |
Revision as of 16:50, 21 September 2010
Ligation
VectorにHeat shock promotorとRBSを組み込む
パーツ | By comparison to marker | size (bp) | (ng) required | (uL) used |
---|---|---|---|---|
Vector | 5 ng/uL | 2996 bp | 10 ng | 2 uL |
Heat shock promotor | 25 ng/uL | 800 bp | 8 ng | 0.3 uL |
RBS | 1 ng/uL | 50 bp | 1 ng | 1 uL |
Total | 3.3 uL |
Ligation and transformation follows same protocol as previously
competencyを求める
- 0.35 ug/uLのpUC119 DNAを1 uL 0.5 mLチューブにとり,2.5 uLのDWを加えた (0.1 ug/uL)
- 999 uLのDWを1.5 mLチューブにとり,1 uLの[1]を加えた (0.1 ng/uL)
- 99 uLのDWを1.5 mLチューブにとり,1 uLの[2]を加えた (1 pg/uL)
1 pg/uLのpUC119 DNAを「青α」と「赤α」のコンピテントセルに,それぞれ1 uL, 10 uL, 100 uLずつ加え,Transformationさせた
VectorだけでLigation,Transformation
前日のLigationでコロニーができなかったので,ベクターに問題があるかもしれない.
- pSB1C3 2 uLにligation solution 2 uLとT4 ligase 0.4 uLを加えた
- Transformation