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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:

#	Ligation reaction
1	3 μL I10341 + 4 μL J1100 + 1.5 μL pSB3C5
2	3 μL I10341 + 4 μL J1101 + 1.5 μL pSB3C5
3	3 μL I10341 + 4 μL J1107 + 1.5 μL pSB3C5
4	3 μL I10341 + 4 μL J1117 + 1.5 μL pSB3C5
5	3 μL I10341 + 4 μL J1127 + 1.5 μL pSB3C5
6	4 μL J1100 + 1.5 μL pSB3C5 (negative control)

We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added. The mixtures were incubated overnight at 16 °C.