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Team:HokkaidoU Japan/Notebook/August19
From 2010.igem.org
since 13 Jul 2010
since 1 Aug 2010
Ligation
Ligated Vector with Heat shock promotor and RBS
| Parts | 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 Check
- Transfered 1 uL of pUC119 DNA (0.35 ug/uL) into 0.5 mL tube and added 2.5 uL of ADW (0.1 ug/uL)
- Took new 1.5 mL tube and added 999 uL of ADW (0.1 ng/uL) to it plus 1 uL of [1]
- Took new 1.5 mL tube and added 99 uL of ADW to it plus 1 uL of [2]
- Added pUC119 DNA (1 pg/uL) to each DH5alpha tube of 50ul and 30ul also varying the amount of pUC119 DNA:1 uL, 10 uL, 100 uL
- Transformed
Ligation and Transformation between Vector，Transformation
Because there weren't any colonies the previous day we suspected that there might be something wrong with vectors
- Mixed 2 uL of pSB1C3, 2 uL of ligation solution and 0.4 uL of T4 ligase
- Transformed
