2.1 Prepare Transposition Mix

  1. Prepare Transposition Mix on ice. Pipette mix 10x and centrifuge briefly.

     

    Transposition Mix

    Add reagents in the order listed

     

    PN

     

    1X (µl)

    4X +

    10% (µl)

    8X +

    10% (µl)

    ATAC Buffer C

    2001581

    5.0

    22.0

    44.0

    ATAC Enzyme B

    2001582/

    2001583

     

    5.0

     

    22.0

     

    44.0
     

    Total

    -

    10.0

    44.0

    88.0

  2. Add 10 μl Transposition Mix to a tube of a PCR 8-tube strip for each sample. Centrifuge briefly and maintain on ice.

  3. Scalable Transposition: Refer to Scalable Transposition – Nuclei Input Calculator to calculate the volume of Nuclei Stock and Diluted Nuclei Buffer for a total volume of 15 μl.

  4. Add the calculated volume of Diluted Nuclei Buffer to the Transposition Mix. Pipette mix 10x (pipette set to 10 μl). Centrifuge briefly.

  5. Gently pipette mix the Nuclei Stock. Add the calculated volume of the Nuclei Stock to the tube containing the Transposition Mix. Gently pipette mix 5x (pipette set to 20 μl). DO NOT centrifuge.