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    EZ DNA Methylation-Startup Kit


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    Cat # Name Size Price Quantity
    D5024 EZ DNA Methylation-Startup Kit 50 Rxns. €455,00
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    Highlights

    • The complete solution for bisulfite conversion. This all-in-one kit contains reagents for bisulfite conversion of DNA without the need for purification, methylated human DNA with control primers, and a robust hot-start PCR polymerase that is specifically formulated for bisulfite-converted DNA.
    • Designed for the first-time user requiring a consolidated product to control for bisulfite conversion.
    • Kit includes: one EZ DNA Methylation-Direct kit, one fully methylated Universal Methylated Human DNA Standard, and ZymoTaq DNA Polymerase.
    Description & Documents Specifications FAQ Components

    Documents

    Product Description

    The EZ DNA Methylation-Startup Kit provides the necessary tools for complete bisulfite conversion of DNA for methylation analysis. This kit includes reagents that allow for bisulfite conversion of purified DNA and DNA directly from blood, cells, and fresh or FFPE tissues without the prerequisite for upstream DNA purification. A fully methylated Universal Methylated Human DNA Standard is provided with a special primer set for PCR to assess conversion efficiency. Finally, a unique ZymoTaq DNA Polymerase is included for robust amplification of bisulfite-treated DNA.

    Documents

    Technical Specifications

    Applicable For Recovered DNA is ideal for downstream analyses including PCR, endonuclease digestion, sequencing, microarrays, etc.
    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 ≥ 10 µl
    Equipment Thermocycler with heated lid and microcentrifuge.
    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.

    Resources

    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.

    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.

    > 50 bp.

    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.

    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.

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

    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).

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

    Cat # Name Size Price
    E2003 ZymoTaq PreMix (50 Rxns.) (2 x 625 µl) €84,20
    D5020 EZ DNA Methylation-Direct Kit 50 Rxns. €242,80
    D5011 Universal Methylated Human DNA Standard Human €217,00

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