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    EZ-96 DNA Methylation-Direct MagPrep


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    Cat # Name Size
    D5045 EZ-96 DNA Methylation-Direct MagPrep 8 x 96 Rxns.
    D5044 EZ-96 DNA Methylation-Direct MagPrep 4 x 96 Rxns.

    Highlights

    • High throughput, complete bisulfite conversion of DNA directly from blood, tissue, cells, FFPE and LCM-derived samples
    • Compatible with small sample inputs – as few as 10 cells or 50 pg of DNA.
    • High throughput (96-well), automated desulphonation and recovery of bisulfite-treated DNA.
    Description & Documents Specifications FAQ Components Citations

    Documents

    Supplemental Info

    Product Description

    The EZ-96 DNA Methylation-Direct MagPrep is a magnetic bead-based bisulfite conversion kit that allows simple and reliable DNA bisulfite conversion directly from blood, tissue, and cells without the prerequisite for DNA purification. The increased sensitivity of this magnetic bead-based bisulfite conversion kit makes it possible to amplify bisulfite-converted DNA from as few as 10 cells or 50 pg DNA. These innovations have been coupled to a magnetic bead-based clean-up for high-throughput methylation analysis. Desulphonation and clean-up of the converted DNA is performed while bound to the MagBinding Beads. This magnetic bead based bisulfite conversion kit has been designed to minimize template degradation, loss of DNA during treatment and clean-up, and to provide complete conversion of unmethylated cytosines. Recovered DNA is ideal for downstream analyses, including PCR amplification, endonuclease digestion, sequencing, microarrays, etc.

    For automation scripts and support, email automation@zymoresearch.com

    Documents

    Technical Specifications

    Applications Purified, converted DNA is of high-quality and well-suited for downstream processes, including library preparation for Next-Generation sequencing, PCR amplification, etc.
    Conversion > 99.5%
    Elution Volume ≥ 25 µl
    Equipment Thermocycler with heated lid, heating element for 96-well plate, magnetic stand.
    Input Samples containing between 50 pg to 2 µg of DNA. For optimal results, the amount of input DNA should be from 200 to 500 ng. The number of cells per standard treatment can range from 10-105 cells.
    Processing Time 4 hours
    Recovery > 80%
    Sample Source Blood, tissue, cells, FFPE, LCM-derived samples, purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc.
    Supplemental Info

    Resources

    https://www.zymoresearch.com/pages/bisulfite-beginner-guide
    https://www.zymoresearch.com/pages/bisulfite-primer-seeker

    Leaving the desulphonation buffer on the column longer than recommended will cause more degradation and result in lower yields.

    Bisulfite conversion will work regardless of context, so the kits are compatible with genomic DNA derived from plants and other species with high non-CpG methylation levels.

    ZymoTaq DNA Polymerase has been specifically designed for use in bisulfite amplification reactions. ZymoTaq is available as a stand-alone polymerase (E2001/E2002), PreMix (E2003/E2004), and qPCR PreMix (E2054/E2055).

    > 50 bp.

    For best results, keep the method of quantification consistent before and after bisulfite treatment:

    • If quantifying with a NanoDrop, use dsDNA settings (50 μg/ml for Ab260 = 1.0) before treatment and use RNA settings (40 μg/ml for Ab260 = 1.0) after treatment.
    • If quantifying with Qubit, use a dsDNA assay before treatment and use a ssDNA assay after treatment.
    Following bisulfite treatment of DNA, nonmethylated cytosine residues are converted into uracil. The recovered DNA is typically A, U, and T-rich. The original base-pairing no longer exists. Instead, the converted DNA is single stranded with limited non-specific base-pairing at room temperature. Therefore, traditional dsDNA methods of quantification will not accurately represent the amount of recovered bisulfite converted DNA.

    Following bisulfite treatment, DNA will be single stranded with limited non-specific base pairing at room temperature. To visualize, run the converted DNA on an agarose gel then chill the gel on ice or in an ice bath for 30 minutes. This will force enough base-pairing to allow intercalation of the ethidium bromide for the DNA to be visible. If using a Bioanalyzer or TapeStation instrument, use RNA kits and reagents to visualize the converted DNA.

    Poor conversion efficiency and low yields can be due to a variety of different experiment-specific conditions. Please contact Technical Support to discuss your specific experimental conditions and further troubleshoot with a product specialist.

    Converted DNA eluted in M-Elution Buffer can generally be stored at -20°C for 1-3 months. If longer term storage is necessary, we recommend storing at or below -70°C if possible. Bisulfite converted DNA is less stable than dsDNA; for best results, minimize freeze-thawing of converted DNA and use as soon as possible for downstream analysis.

    Cat # Name Size
    D4100-5-8 MagBinding Beads 8 mL
    D4100-5-16 MagBinding Beads 16 mL
    C2003 Elution Plate 2 Plates
    C2002 Collection Plate 2 Plates
    C2005 Conversion Plate w/ Cover Foil 2 Plates/Foils
    D5021-7 M-Solubilization Buffer 18 ml
    D5021-8 M-Reaction Buffer 4ml
    D5021-9 M-Digestion Buffer (2X) 15 ml
    D5040-3 M-Binding Buffer 250 ml
    D5040-5 M-Desulphonation Buffer 80 ml
    D5040-4 M-Wash Buffer 72 ml
    D5007-6 M-Elution Buffer 8 ml
    D5006-2 M-Dilution Buffer-Gold 7 ml
    D5003-1 CT Conversion Reagent (96 Conversions) 1 Bottle

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