Home FAQs 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 Blunt/TA Ligase Master Mix my transformation plates have very few colonies. What do you think is the problem?

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 Blunt/TA 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 Blunt/TA 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 a Blunt/TA 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 Blunt/TA Ligase Master Mix. It is not necessary to use more than 5 ul in a standard transformation reaction.