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    EZ-96 DNA Methylation MagPrep

    Highlights

    • Proven procedure for complete bisulfite conversion of DNA.
    • High-throughput (96-well), automated desulphonation and recovery of bisulfite-treated DNA.
    • Recovered DNA is ideal for downstream analyses including PCR, endonuclease digestion, sequencing, microarrays, etc.
    Zymo Research's 100% Satisfaction Guarantee

    Original Manufacturer

    Innovated in California, Made in the USA

    Satisfaction 100% guaranteed, read Our Promise

    Highlights

    • Proven procedure for complete bisulfite conversion of DNA.
    • High-throughput (96-well), automated desulphonation and recovery of bisulfite-treated DNA.
    • Recovered DNA is ideal for downstream analyses including PCR, endonuclease digestion, sequencing, microarrays, etc.
    Zymo Research's 100% Satisfaction Guarantee

    Original Manufacturer

    Innovated in California, Made in the USA

    Satisfaction 100% guaranteed, read Our Promise

    Cat # Name Size
    D5040 EZ-96 DNA Methylation MagPrep 4 x 96 Rxns.
    D5041 EZ-96 DNA Methylation MagPrep 8 x 96 Rxns.
    Cat # Name Size
    C2003 Elution Plate 2 Plates
    C2002 Collection Plate 2 Plates
    C2005 Conversion Plate w/ Cover Foil 2 Plates/Foils
    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
    D5003-1 CT Conversion Reagent (96 Conversions) 1 Bottle
    D5041-6 M-Elution Buffer 40 ml
    D5002-2 M-Dilution Buffer 5.2 ml
    D4100-5-8 MagBinding Beads 8 mL
    D4100-5-16 MagBinding Beads 16 mL

    Description

    The EZ-96 DNA Methylation MagPrep is a bisulfite conversion kit that features a high-throughput (96-well), simplified procedure that streamlines bisulfite conversion of DNA. The kit is based on the three-step reaction that takes place between cytosine and sodium bisulfite during which cytosine is converted into uracil. Desulphonation and clean-up of the converted DNA is performed while bound to the MagBinding Beads. The kit is designed to reduce template degradation and minimize DNA loss during treatment and clean-up, while ensuring complete conversion of the DNA. Purified, converted DNA is ideal for downstream analyses including PCR amplification, endonuclease digestion, sequencing, microarrays, etc.

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

    Performance

    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%
    Equipment Thermocycler with heated lid, heating element for 96-well plate, magnetic stand.
    Input 500 pg - 2 µg of DNA
    Processing Time 12-16 hours
    Recovery > 70%
    Sample Source Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.

    Resources

    Documents

    FAQ

    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).

    Citations

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