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Team:Cambridge/Notebook/12
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Week 11: Monday 27th September - Sunday 3rd October
113. Expt: Biobrick assembly of fluorescent proteins (YFP, CFP, RFP) into PP+pBAD+pSB1C3 and pBAD+pSB1C3 (Emily and Bill)
- Miniprepped overnight cultures of YFP, CFP, RFP with rbs's and PP+pBAD+pSB1C3
- Nanodrop readings:
| Nanodrop reading (ng/µl) | |
| Cm+PP+pBAD+pSB1C3 | 48.5 |
| YFP+rbs | 74.8 |
| RFP+rbs | 32.9 |
| CFP+rbs | 46.7 |
pSB1C3 from freezer was used at 12.9ng/µl
- Restrict using protocol on p88, using following quantities:
| Cm+PP+pSB1C3 | YFP (for PP) | YFP (for pBAD) | RFP (for PP) | RFP (for pBAD) | CFP (for PP) | CFP (for pBAD) | pSB1C3 | |
| Nuclease-free H20 | 1 | 6 | 6 | 1 | 1 | 6 | 6 | 1 |
| 10x FD Buffer | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 |
| Plasmid DNA | 15 | 10 | 10 | 15 | 15 | 10 | 10 | 16 |
| EcoRI | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
| SpeI | 0 | 1 | 0 | 1 | 0 | 1 | 0 | 0 |
| XbaI | 1 | 0 | 1 | 0 | 1 | 0 | 1 | 0 |
| PstI | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
| * | * | * | * |
µl°C
