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    EZ DNA Methylation-Lightning Kit

    D5030-E / D5031-E / D5030T / D5030 / D5031


    EZ DNA Methylation-Lightning Kit

    Cat # Name Size
    D5030-E EZ DNA Methylation-Lightning Kit (CE-IVD) 50 Rxns.
    D5031-E EZ DNA Methylation-Lightning Kit (CE-IVD) 200 Rxns.
    D5030T EZ DNA Methylation-Lightning Kit 10 Rxns.
    D5030 EZ DNA Methylation-Lightning Kit 50 Rxns.
    D5031 EZ DNA Methylation-Lightning Kit 200 Rxns.

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    Highlights

    • Fastest bisulfite conversion kit for complete bisulfite conversion of DNA for methylation analysis.
    • Ready-to-use conversion reagent is added directly to DNA.
    • High-yield, converted DNA is ideal for PCR, Methylation Specific PCR (MSP), arrays, library preps, Next-Generation sequencing, etc.
    Description

    The EZ DNA Methylation-Lightning Kit can rapidly bisulfite convert and purify DNA in less than 1.5 hours. A ready-to-use conversion reagent streamlines this process – simply add the reagent directly to the sample and incubate. Desulphonation and clean-up of the converted DNA is performed on a spin-column, allowing an elution volume as low as 10 µl. The kit achieves high conversion efficiency of unmodified cytosines into uracil, ensuring accurate downstream methylation analysis.


    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 100 pg - 2 µg of DNA.
    Processing Time 1.5 hours
    Recovery > 80%
    Sample Source Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.
    Supplemental Info

    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).

    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.

    > 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.




    “Results were as good as with the formerly used EZ DNA Methylation Gold Kit but significantly reduced total time for analyses. More flexibility due to the possibility of up to 20h storage of the samples after bisulfite conversion before the cleaning procedure especially for part-time employees.”

    - Sabrina S. (UK-Aachen)

    “The conversion reagent no longer has to be made up from individual components and dissolved - it is supplied ready-made. The conversion reagent can be stored at room temperature - there is no need to use up all 10 uses in one month. This makes it more flexible, and avoids wastage. Shorter protocol saves time.”

    - Jeremy B. (AgResearch)


    Cat # Name Size
    C1004-50 Zymo-Spin IC Columns 50 Pack
    C1001-50 Collection Tubes 50 Pack
    D5002-4 M-Wash Buffer 24 ml
    D5001-4 M-Wash Buffer 6 ml
    D5005-3 M-Binding Buffer 30 ml
    D5002-6 M-Elution Buffer 4 ml
    D5001-6 M-Elution Buffer 1 ml
    D5006-3 M-Binding Buffer 125 ml
    D5030-5 L-Desulphonation Buffer 10 ml
    D5031-5 L-Desulphonation Buffer 40 ml
    D5030-1 Lightning Conversion Reagent 1.5 ml