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    PureRec Duplex-Specific Nuclease (DSN)

    E1020-200 / E1020-1000

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    PureRec Duplex-Specific Nuclease (DSN)

    Get Free Sample
    Cat # Name Size Price Quantity
    E1020-200 PureRec DSN 200 U $148.00
    - +
    E1020-1000 PureRec DSN 1000 U $518.00
    - +

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    Highlights
    • Highest Specificity: Selectively digests dsDNA and DNA in DNA-RNA hybrids; virtually inactive towards ssDNA or RNA.
    • Premium Purity: RNase-free, protease-free, and animal-free.
    • Thermostable: Able to withstand high temperatures without denaturation or loss in activity.
    Description

    PureRec Duplex-Specific Nuclease (DSN) is an endonuclease that selectively digests double-stranded DNA (dsDNA) and DNA in DNA-RNA hybrids, exhibiting virtually no activity towards single-stranded DNA (ssDNA) or RNA.

    It can discriminate between perfectly matched and non-matched short DNA duplexes. To catalyze cleavage, DSN needs a minimum of 10 bp dsDNA or 15 bp perfectly matched DNA-RNA duplex.

    This enzymatic function is reliant upon the presence of Mg2+ ions, with a recommended concentration of 5 mM for most applications.

    PureRec DSN is overexpressed and extensively purified from a non-animal source and is free of non-specific nucleases or RNases.

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    Application
    • Remove gDNA and DNA in DNA:RNA hybrids for protein or RNA preparations.
    • Reduce abundant rRNA and globin transcripts to enhance next-generation sequencing (NGS) libraries.
    • Achieve precise single-nucleotide polymorphism (SNP) detection with minimal off-target effects.
    • Enhance signal amplification for accurate virus detection.
    • Improve detection of miRNA and biomarkers.
    Unit Definition One Kunitz unit causes an increase in absorbance at 260 nm of 0.001 per minute per ml, at 25°C, pH 7.15, when acting on calf thymus DNA.
    Concentration 1 U/µl
    Optimal Temperature 60 °C
    Reaction Buffer 10X DSN Digestion Buffer
    500 mM Tris-HCl, 50 mM MgCl2, pH 8.0
    Stop Solution 2X DSN Stop Solution
    10 mM EDTA, pH 8.0
    Storage Condition For long-term storage, store at -20 °C.
    Inactivation Add the 2X DSN Stop Solution to the reaction, to a final concentration of 1X, and incubate at the reaction temperature for 5 minutes.

    • This depends on the application. DSN is a highly versatile enzyme, and the input amount can be adjusted to meet specific requirements. The amount of DSN and incubation times will vary based on the desired product. For example, a typical amount of DSN used for the normalization of cDNA libraries ranges from 0.25 to 1 U.
    • To protect ssDNA in a pool of dsDNA/ssDNA, we recommend titrating the enzyme amount to determine the optimal conditions for ssDNA protection.

    The incubation temperature can be adjusted between 25°C and 70°C, with an optimal temperature of 60°C.

    DSN is susceptible to EDTA inactivation at concentrations of 5 mM and above. We recommend performing an inactivation step by adding 5 mM EDTA at the reaction temperature for 5 minutes following digestion.

    DSN is thermally stable and retains activity when heated to 98°C for 10 minutes. We suggest stopping all reactions with DSN by incubating with 5 mM EDTA for 5 minutes at 60°C.

    Yes, we can customize your order request to meet your specific requirements. For bulk ordering, please fill out the form and one of our specialists will be in touch shortly.

    Cat # Name Size Price
    E1020-4-50 2X DSN Stop Solution 50 mL $28.00
    E1020-2-5 10X DSN Digestion Buffer 5 mL $19.00

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