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

  1. Add 1 μg in up to 200 μl input DNA to the tube containing the 160 μl of MBD2-Fc-bound magnetic beads. For other DNA input amounts, add DNA to (Y) μl of MBD2-Fc bound magnetic beads.

    Note: Y is calculated here.

  2. Add undiluted Bind/wash Buffer (5X) for a final concentration of 1X. (For example add 10 μl of Bind/wash Buffer (5X) if the DNA input sample was 40 μl, add 4 μl of Bind/wash Buffer (5X) if the DNA sample was 16 μl). Pipette the sample up and down until the beads are completely homogenous, at least 5-10 times.

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

  3. Agitate the tube on a rotating mixer for 15 minutes at room temperature with rotation.