Capture Methylated Host DNA - NEBNext Microbiome DNA Enrichment Kit (E2612)

  1. Add up to 1 μg input DNA to the tube containing the 160 μl of MBD2-Fc-bound magnetic beads. For other DNA input sizes, add DNA to (Y) μl of MBD2-Fc bound magnetic beads.
    Note: Y is calculated here.

  2. Add Bind/wash Buffer (5X) in the ratio determined by the volume of the DNA input sample (from previous step) for a final concentration of 1X (e.g, add 10 μl of Bind/wash Buffer (5X) if the DNA input sample was 40 μl, 4 μl of Bind/wash Buffer (5X) if the DNA sample was 16 μl). Pipet the sample up and down a few times to mix.

    Volume of 5X Bind/wash Buffer to add (μl) = Vol input DNA (μl)/4

  3. Incubate the reaction for 15 minutes at room temperature with rotation.