Team:TU Delft/23 June 2010 content

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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.