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  • FAQ: I routinely use more than 5 ul of my ligation reactions to transform 50 ul aliquots of competent cells. When I do this with the Instant Sticky-end Ligase Master Mix my transformation plates have very few colonies. What do you think is the problem?

    We do not recommend the use of more than 5 ul of Instant Sticky-end Ligase Master Mix ligation reactions to transform 50 ul of competent cells. In some instances we have observed a decrease in the transformation efficiency when using more than 5 ul of an Instant Sticky-end Ligase Master Mix ligation reaction. DNA with compatible ends that are intact and present in the amounts specified by the protocol should be ligated by the Instant Sticky-end Ligase Master Mix. It is not necessary to use more than 5 ul in a standard transformation reaction.