Team:HokkaidoU Japan/Notebook/August24
From 2010.igem.org
(Difference between revisions)
(→ラオリプライマーで増やしたパーツの確認) |
(→Ligation Mixtureのチェック) |
||
Line 29: | Line 29: | ||
* This vector didn't have a primer annealing site for our new primers | * This vector didn't have a primer annealing site for our new primers | ||
- | =Ligation | + | =Check of Ligation Mixtures= |
- | Ligation | + | TAKARABIO's Ligation Mixture [I] and Ligation Mixture Mighty Mix [M] |
- | * | + | * After PCR and gel extracted pSB1C3 volume of either Ligation Mixture equal to pSB1C3 solution volume was added |
- | * | + | * Transformed |
+ | * incubated in 200 uL of LB medium | ||
+ | * 100 uL of each solution was plated on mediums containing 34 ug/uL and 12 ug/uL of chloramphenicol | ||
=いろいろ確認= | =いろいろ確認= |
Revision as of 08:52, 22 September 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
Check of Ligation Mixtures
TAKARABIO's Ligation Mixture [I] and Ligation Mixture Mighty Mix [M]
- After PCR and gel extracted pSB1C3 volume of either Ligation Mixture equal to pSB1C3 solution volume was added
- Transformed
- incubated in 200 uL of LB medium
- 100 uL of each solution was plated on mediums containing 34 ug/uL and 12 ug/uL of chloramphenicol
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