TSAP Thermosensitive Alkaline Phosphatase (100 units)

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.

For more information, see the Protocols & Applications Guide.

Notes

Units listed are MBU (molecular biology units).

References

1. Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.