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Team:HokkaidoU Japan/Notebook/aggregation Week 6
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Revision as of 06:20, 6 August 2012
August 6th
Ethanol precipitation
Ethanol precipitation for digestion and gel extraction product.
- Added 5 ul of NaoAc, 1.5 ul of glycogen and 125ul of 100% ethanol.
- Centrifuged in 15000 rpm, 10 min at 4C.
- Remove supernatant and added 220ul of 70% ethanol.
- Centrifuged in 15000 rpm, 10 min at 4C.
- Remove supernatant and air drying in room temperature then added 10 ul of DW.
Ligation
We ligated pT7-RBS on pSB1K3 (digested Spe1) as vector and Ag43-dT (digested Spe1 and Xba1 and HindIII) as insert.
| pT7-RBS | 1 ul |
| Ag43-dT | 2 ul |
| DW | 2 ul |
| Ligation Mighty Mix(TAKARA) | 5 ul |
| Total | 10 ul |
Ligation reaction time was written below.
| Degree | Minute |
| 16 | 30 |
| 65 | 10 |
| 4 | Hold |
