8.3 Adaptor Ligation

  1. Prepare Adaptor Ligation Mix. Pipette mix and centrifuge briefly.

    2

     

    Fixed percentage added to total to help with pipetting

    Adaptor Ligation Mix
    Add reagents in the order listed

    PN

    Volume for Master Mix (μl)

    Scaled Volume (μl)

    Ligation Buffer

    2001109/2001317

    40

     

    DNA Ligase

    		 220110/
    220131/2001318    		 10

     

    Total

    		 

    50

     

  2. Add 50 μl Adaptor Ligation Mix to 50 μl sample. Pipette mix 15x (pipette set to 90 μl). Centrifuge briefly.

  3. Incubate in a thermal cycler with the following protocol.

    Lid Temperature

    Reaction Volume

    Run Time

    hh:mm:ss

    30°C

    100 µl

    15 min

    Step

    Temperature

    Time

    1

    20°C

    00:15:00

    2

    4°C

    Hold