TSAP Thermosensitive Alkaline Phosphatase
TSAP Thermosensitive Alkaline Phosphatase catalyzes the removal of 5' phosphate groups from DNA, thus preventing the recircularization and religation of linearized cloning vector DNA during ligation. It is effective on 3' overhangs, 5' overhangs and blunt ends. It is also useful for preparing DNA for 5' end-labeling by removing existing phosphate groups from the 5' end. TSAP is irreversibly inactivated by heating at 74 degrees C for 15 minutes. Therefore, a DNA cleanup step is not required before proceeding to a ligation reaction. TSAP is fully active in all restriction enzyme reaction buffers tested under the conditions listed below, facilitating a streamlined restriction digestion, dephosphorylation and ligation reaction.