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    EZ DNA Methylation-Lightning Automation Kit

    D5049


    EZ DNA Methylation-Lightning Automation Kit

    Cat # Name Size
    D5049 EZ DNA Methylation-Lightning Automation Kit 96 preps

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    Highlights

    • Automation-specific streamlined design for high-throughput bisulfite conversion of DNA for methylation analysis.
    • Compatible with Illumina® Infinium™ MethylationEPIC, HTS iSelect® Methyl Custom, and Mouse Methylation arrays.
    • Optimized buffer volumes for consistent automation performance.
    Description

    The EZ DNA Methylation-Lightning Automation Kit is a high-throughput DNA bisulfite conversion kit designed for streamlining automation workflows. It is recommended for several downstream Illumina® Infinium® products such as MethylationEPIC, HTS iSelect® Methyl Custom, and Mouse Methylation BeadChip Arrays. This automation-specific kit features optimized buffer volumes for consistent performance on all open platform liquid handlers and magnetic bead transfer automated systems. Verified automation scripts are readily available for use on the Hamilton Microlab® STAR™/STARlet™, Thermo Fisher Scientific KingFisher™ Flex, and Tecan Fluent®.

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


    Binding Capacity 1 µg total DNA per 5 µl MagBinding Beads
    Conversion Efficiency > 99.5% of non-methylated C residues are converted to U; > 99.5% protection of methylated cytosines.
    DNA Purity A260/A280 & A260/A230 ≥ 1.8
    Input 100 pg - 2 µg of DNA. For optimal results, the amount of input DNA should be from 200 to 500 ng.
    Input Recovery > 80% (≥ 100 bp)
    Required Equipment Thermal cycler with heated lid.
    Sample Source Purified genomic DNA, endonuclease-digested DNA, linearized plasmid DNA, etc. DNA should be high-quality and RNA-free.
    Supplemental Info
    User Provided Materials Ethanol (95 – 100%), non-skirted 96-well microplates, and 96-well microplate sealing films.

    See Recommendations for instrument hardware and labware requirements.

    The EZ DNA Methylation-Lightning automation kit features optimized buffer volumes that allow for consistent high-throughput automation performance.

    While we do not offer scripts for non-verified instruments, we can provide general technical support for any automation instrument.

    Yes, we provide a guideline for users to create their own script in the protocol.

    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.

    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
    D5032-1 Lightning Conversion Reagent 15 ml
    D5049-3 M-Binding Buffer 100 ml
    D5007-4 M-Wash Buffer 36 ml
    D5046-5 L-Desulphonation Buffer 80 ml
    D5049-6 M-Elution Buffer 50 ml