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From 2010.igem.org

LAB WORK

Ligations were performed using the digested BioBricks obtained in the early morning. The ligation protocol was in accordance with the following:

-	μL
I10341	3
J611XX	4
pSB3C5	1.5
Ligation Buffer	3
T4 ligase	1 unit
H20	15

We adhered to the 3:1 insert:plasmid ratio when determining the volumes of DNA added. The negative control for the ligation consisted of J61100 + pSB3C5. The mixtures were incubated overnight at 16 degrees.