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Alkaline phosphatase

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    Product description
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    Product Description
    Alkaline Phosphatase, the Chinese name alkaline phosphatase, can remove the 5'end phosphate groups of DNA and RNA. It is often used to prevent the self-ligation of vectors: in molecular cloning experiments, DNA ligase requires a phosphate group to catalyze DNA ligation In the presence of the group, the carrier will retain a phosphate group at the end of the cleavage point after the carrier is cleaved. After the carrier is dephosphorylated by alkaline phosphatase, there is no phosphate group at the 5'end, so it cannot be connected to its 3'end. Therefore, the preferential ligation reaction of the vector itself in the ligation reaction is prevented, which improves the insertion rate of the target fragment. In addition, alkaline phosphatase can also prepare DNA templates for 5'end labeling, and for dephosphorylation of the enzyme protein.
    This product is alkaline phosphatase derived from calf intestine, which can degrade almost all phosphate monoesters, but this enzyme cannot hydrolyze phosphodiester or phosphotriester.
    Source: Recombinant Escherichia coli (calf intestine ALP gene).
    Specification: 30 U/µL.
    Application
    1. Dephosphorylation of cloning vector DNA to prevent recirculation during ligation;
    2. Degradation of dNTPS before PCR product sequencing;
    3. Dephosphorylation of the 5'-end of nucleic acid before labeling with T4 polynucleotide kinase;
    4. Other applications for dephosphorylation of DNA and RNA substrates.
    Transportation and storage methods
    Ice pack transportation. Store the product at -20°C.
    Activity definition
    At 37°C and pH 9.8, the amount of alkaline phosphatase used to generate 1 µmol of nitrophenol in 1 minute with p-nitrophenol phosphate as the substrate is defined as 1 activity unit (U).
    Inactivation
    In the presence of a chelating agent, heating at 65°C for 30 minutes will cause 99% of the activity to be irreversibly lost; the use of phenol can completely inactivate the enzyme.
    Quality Inspection
    No endo or exonuclease, ribonuclease contamination.
    Storage buffer
    100 mM Tris-HCl (pH 8.0), 1 mM MgCl2, 0.1 mM ZnCl2, 0.1% (v/v) Triton X-100 and 50% (v/v) glycerol.
    10 X ALP Buffer
    1 M Tris-HCl (pH 8.0, 37°C), 100 mM MgCl2, 0.1 mM ZnCl2, 100 mM KCl.
    Optimal pH
    When the substrate concentration is high, the optimal pH of the enzyme is 10; when the substrate concentration is low, the optimal pH is 8.
    Precautions
    1. For your safety and health, please wear lab coats and disposable gloves for operation.
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