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Team:Chiba/Notebook/Calendar/29 September 2009
From 2009.igem.org
(28_September_2009 <|>30_September_2009)
Contents |
Gel electrophoresis
Yesterday's oparation is here.
- Today's operation
10:40-
Mini Prep.
13:30-
Gel electrophoresis(135 V, 27 min)
---DNA Clean & Concentrator---
15:50-
Gel electrophoresis again(135 V, 27 min)
Digestion Test
- Sample
Vector(plux-GFP)
Insert(sfGFP)
- Vector
| Vector DNA | 30 μL |
| Nco1 | 1 μL |
| BamH1 | 1 μL |
| Buffer3 | 5 μL |
| BSA | 5 μL |
| NFW | 8 μL |
| Total | 50 μL |
- Insert
| Insert DNA | 20 μL |
| Nco1 | 1 μL |
| BamH1 | 1 μL |
| Buffer3 | 3 μL |
| BSA | 3 μL |
| NFW | 2 μL |
| Total | 30 μL |
18:55-
Gel electrophoresis(135 V, 25 min)
19:45
Took a picture
Transformation
- Sample
Competent Cells : XL10G(K)
Plasmid : plux-GFP(Cm)
21:45-
CmのプレートにまいてCultured it at 37 degrees Celsius.
Digestion
- Sample
Vector : plux-GFP(p15A)
Insert : sfGFP
Enzymes : Nco1, BamH1
21:17
We added Enzymes and took it 37 degrees Celsius.
---3 h---
We took it 65 degrees Celsius for 酵素を失活させる
---20 min---
24:42
We kept it in refrigerator.
PCR
- Template
plux-HSUTK
- Element of mixtures
| Template | 1 μL |
| Primer(F) | 2.5 μL |
| Primer(R) | 2.5 μL |
| Buffer(x 10) | 5 μL |
| dNTP | 4 μL |
| DNA pol. | 1 μL |
| dH2O | 34 μL |
| Total | 50 μL |
