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Team:HokkaidoU Japan/Notebook/August17
From 2010.igem.org
since 13 Jul 2010
since 1 Aug 2010
Gel Extraction
Electrophoresis
20 uL of DNA digested yesterday was electrophoresed with 4 ul of 6x SB. All acording to the table bellow.
| Lane | DNA |
| 1 | Empty |
| 2 | λ/Hind III |
| 3 | Empty |
| 4 | pSB1C3/E, P |
| 5 | pSB1C3/E, P |
| 6 | Heat Shock Promotor/E, S |
| 7 | Heat Shock Promotor/E, S |
| 8 | RBS/X, P |
| 9 | RBS/X, P |
| 10 | RFP/E, S |
| 11 | RFP/E, S |
| 12 | double Terminator/E, S |
| 13 | double Terminator/E, S |
| 14 | Empty |
| 15 | λ/Hind III |
| 16 | Empty |
| 17 | Empty |
→Gel Extraction
- After ectraction DNA was electrophoresed to check consentration
- 2 uL of 6x SB was added to DNA solution of 10 uL
| Lane | DNA |
| 1 | Empty |
| 2 | λ/Hind III |
| 3 | Vector |
| 4 | Heat shock promotor |
| 5 | RBS |
| 6 | RFP |
| 7 | double terminator |
| 8 | Empty |
- Band of RBS isn't visible
- This Might due to loss durring extraction
→will try again tomorrow
Making of Chloramphenicol LB Medium
Because we were planing to use pSB1C3 we made LB Medium with chloramphenicol
- Added 12.5 g LB-broth and 7.5 g of Agar 7.5 g to 500 mL to autoclaved distiled water, autoclave
- Added 500 uL of Chloramphenicol (34 mg/mL)
- Poured it to 25 plates, 20 mL per plate
