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Description:
SuperSAP™ is an enhanced formulation of Shrimp Alkaline Phosphatase designed for rapid dephosphorylation of DNA prior to its use in cloning applications and end-labeling. SuperSAP dephosphorylates up to 5 pmols of cohesive or blunt ends with a background reduction  95% in just 5 min. Background reduction is the inhibition of recircularization in a self-ligation reaction and measured by transformation in E. coli. SuperSAP may also be used to dephosphorylate RNA and protein.
Benefits
SuperSAP has all the benefits of SAP. It has a high specific activity and is heat-labile. SuperSAP can be easily heat-inactivated at 65°C for 15 min and works well in various restriction enzyme buffers. No additional buffers or additives are required.
Rapid Dephosphorylation
SuperSAP removes the terminal 5’ phosphate from protruding-, blunt- or recessed-ends in just 5 minutes.
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| Rapid SuperSAP™ dephosphorylation for 5 min prevents recirculation of vector. pUC19 vector was digested as indicated, purified using a spin column and re-suspended in 10mM Tris-HCl, pH 8.5. 5 μg was treated with 1 μl of SuperSAP and incubated for 5 min at 37°C followed by heat-inactivation at 65°C for 15 min. 50 ng was self-ligated directly using the USB Ligate-IT Rapid Ligation Kit (PN 78400), 2.5 ng was transformed into E. coli DH5-a and 0.5 ng was plated on selective medium. |
Easy Vector Preparation for Cloning
SuperSAP works directly in most restriction enzyme digests and can be added directly to the reaction to simultaneously cut and dephosphorylate DNA. No additional buffers or additives are required.
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Simultaneous dephosphorylation with SuperSAP™ and restriction enzyme digestion is convenient and effective to prevent recirculation of vector. 1 μg of pUC19 was digested as indicated with 1 μl of SuperSAP (5' overhang or blunt), or 2 μl of SuperSAP (3' overhang) in the appropriate restriction enzyme buffer and was incubated for 30 min at 37°C followed by heat-inactivation at 65°C for 15 min. 50 ng was directly self-ligated using the USB Ligate-IT Rapid Ligation Kit (PN 78400), 2.5 ng was transformed into E. coli DH5- and 0.5 ng was plated on selective medium. |
Convenient – Direct Ligation
SuperSAP reactions do not require purification, e.g. phenol extraction, ethanol precipitation or spin column, prior to ligation. After SuperSAP treatment, the dephosphorylated cut vector may be directly ligated using the USB Ligate-IT™ Rapid Ligation Kit [PN 78400/10] or T4 DNA Ligase [PN 70005].
Components:
50 Reaction
Pack Size
SuperSAP 50 µl
10X SuperSAP Reaction Buffer 1 ml
SuperSAP is in 50% glycerol and should be stored at -20°C in a non frost-free freezer.
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