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    EZ DNA Methylation-Gold Kit

    D5006 / D5005


    EZ DNA Methylation-Gold Kit

    Cat # Name Size Price Quantity
    D5006 EZ DNA Methylation-Gold Kit 200 Rxns $614.30
    - +
    D5005 EZ DNA Methylation-Gold Kit 50 Rxns. $182.90
    - +

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    Highlights

    • Complete bisulfite conversion of DNA in less than 3 hours.
    • Desulphonation and recovery of bisulfite-treated DNA with a spin column.
    • Recovered DNA is ideal for downstream analyses such as PCR, endonuclease digestion, sequencing, microarrays, etc.
    Description

    The EZ DNA Methylation-Gold Kit is a refinement of our popular EZ DNA Methylation kit and uses heat denaturation instead of chemical denaturation of the input DNA. This allows for the denaturation and bisulfite conversion steps to be consolidated into one step, leading to a much-reduced incubation time. Recovered bisulfite-converted DNA is ideal for PCR amplification for downstream analyses including 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%
    Elution Volume ≥ 10 µl
    Equipment Microcentrifuge and thermocycler with heated lid
    Input 500 pg - 2 µg of DNA.
    Processing Time 3 hours
    Recovery >75%
    Sample Source Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.
    Supplemental Info

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



    “It makes gene specific DNA methylation analysis very simple and anyone with modest technical background can easily perform the experiment using this kit.”

    - Murali B. (Centre for DNA Fingerprinting and Diagnostics)

    “This protocol allows for bisulfite conversion in about 3 hours. This fast conversion is particularly relevant to our experiments, which allows us to PCR amplify and clone the products in 1 day, saving time without loss of quality.”

    - Joseph M. (Sanford-Burnham Medical Research Institute)


    Cat # Name Size Price
    D5002-4 M-Wash Buffer 24 ml $30.90
    D5001-4 M-Wash Buffer 6 ml $12.40
    D5001-5 M-Desulphonation Buffer 10 ml $18.50
    D5002-5 M-Desulphonation Buffer 40 ml $51.90
    D5005-2 M-Dilution Buffer-Gold 1.5 ml $12.40
    D5006-2 M-Dilution Buffer-Gold 7 ml $12.40
    D5005-3 M-Binding Buffer 30 ml $24.70
    D5002-6 M-Elution Buffer 4 ml $12.40
    D5001-6 M-Elution Buffer 1 ml $12.40
    D5006-3 M-Binding Buffer 125 ml $71.70
    D5005-6 M-Dissolving Buffer 500 µl $12.40
    D5006-6 M-Dissolving Buffer 1.2 ml $18.50
    C1001-50 Collection Tubes 50 Pack $17.00
    C1004-50 Zymo-Spin IC Columns 50 Pack $60.00
    D5001-1 CT Conversion Reagent 1 tube for 10 Conversions $12.40

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