Nucleoside Digestion Mix

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The Nucleoside Digestion Mix is an optimized mixture of enzymes that provides a convenient one-step method to generate single nucleosides from DNA or RNA for quantitative analysis by liquid chromatography-mass spectrometry (LC-MS), eliminating the need for sequential multi-step, time-consuming digestion protocols.

  • Convenient one-step protocol
  • Digests both DNA and RNA to single nucleosides
  • Low-glycerol formulation significantly reduces glycerol-induced ion suppression during MS analysis
  • See what others are saying about the Nucleoside Digestion Mix

Ordering Information

M0649S_pdp_800X450
  • 50 reactions
    $115.00
  • Product Information

    The Nucleoside Digestion Mix is a mixture of enzymes that provides a convenient one-step method to generate single nucleosides from DNA or RNA. Optimized for quantitative analysis by liquid chromatography-mass spectrometry (LC-MS), this reagent eliminates the need for sequential multi-step, time-consuming digestion protocols. The Nucleoside Digestion Mix digests ssDNA, dsDNA, DNA/RNA hybrids and RNA (except mRNA cap structures) containing epigenetically modified (m5C, hm5C, f5C, ca5C, m4C, m6A, etc.), unnatural, or damaged bases. Moreover, the low-glycerol formulation (<1%) significantly reduces glycerol-induced ion suppression during mass spectrometry analysis.

    Supplied in: 20 mM Tris-HCl (pH 7.5), 1 mM MgCl2, 2 mM CaCl2, 2 mM ZnCl2, 50 mM NaCl and 0.6% glycerol.



    Figure 1. Global nucleoside analysis of HeLa DNA following incubation with the Nucleoside Digestion Mix

    Representative HPLC chromatogram of individual deoxyribonucleosides obtained from incubation of 1 µg of purified genomic HeLa DNA digested with 1 µl of the Nucleoside Digestion Mix for 1 hour at 37°C. Deoxyribonucleosides were separated by reversed-phase HPLC and detected by UV absorbance at 260 nm.


    Figure 2. Global nucleoside analysis of HEK 293 RNA following incubation with the Nucleoside Digestion Mix


    Representative HPLC chromatogram of individual ribonucleosides obtained from incubation of 1 µg of purified genomic HEK 293 RNA digested with 1 µl of the Nucleoside Digestion Mix overnight at 37°C. Ribonucleosides were separated by reversed-phase HPLC and detected by UV absorbance at 260 nm. The insert shows an expansion of the chromatogram highlighting the detection of modified ribonucleosides.


    Figure 3. The activity of the Nucleoside Digestion Mix remains stable even after 50 freeze-thaws


    After undergoing multiple freeze-thaw cycles, 1 µl of the Nucleoside Digestion Mix was used to digest 1 µg of λ DNA (NEB #N3011) for 1 hour at 37°C. The nucleoside content of the digested samples was analyzed by LC-MS.

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    Nucleoside Digestion Mix Reaction Buffer -20 10 X
    Product Categories:
    Epigenetics,
    Epigenetic Analysis (EpiMark® Validated),
    DNA Modifying Enzymes
    ,
    Nucleases
    Applications:
    RNA Analysis,
    RNA Modification,
    Methylated DNA Analysis Products
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