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Team:TU Delft/23 June 2010 content
From 2010.igem.org
Lab work
Characterization of Anderson RBS sequences
Ligations were performed using the overnight digested BioBricks. The following ligation reactions was performed:
1. I10341 + J1100 + pSB3C5
2. I10341 + J1101 + pSB3C5
3. I10341 + J1107 + pSB3C5
4. I10341 + J1117 + pSB3C5
5. I10341 + J1127 + pSB3C5
6. J1100 + pSB3C5 (negative control)
The general composition was:
| Component | Sample |
| I10341 | 3 μL |
| J611XX | 4 μL |
| pSB3C5 | 1.5 μL |
| Ligation Buffer | 3 μL |
| T4 ligase | 1 μL |
| H20 | 15 μL |
We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added.
The mixtures were incubated overnight at 16 °C.
