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°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.
Features - Benefits
Easy To Use: TSAP is active in all Promega restriction enzyme buffers, eliminating any cleanup steps or buffer swaps.
Convenient: TSAP is irreversibly inactivated by heating at 74°C for 15 minutes. This allows streamlining of the restriction enzyme digestion, dephosphorylation and ligation procedure by eliminating the need for cleanup after alkaline phosphatase treatment.
Blue/White Cloning-Qualified: Promega's blue/white cloning assay provides a higher level of quality control for enzymes used in cloning applications.
Provided with Promega MULTI-CORE™ Buffer.
Applications
Preventing religation of linearized cloning vehicle DNA by removing phosphate groups from both 5´ termini.
Removing 5´ phosphate groups prior to end-labeling with T4 Polynucleotide Kinase.
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