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Ligations
Ligation reactions between vectors and inserts were set up by adding 1l of freshly
prepared 7.5 mM ATP, 1.5 l of T4 DNA Ligase buffer, insert and vector samples (combined
volume of 11 l) and lastly 1.5 l of T4 DNA Ligase, to give a total volume of 15 l. Reaction
mixtures were incubated at 12oC for at least 16 hours.