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  • Adaptor Ligation (E7370)

     This caution sign signifies a step in the protocol that has multiple paths leading to the same end point but is dependent on a user variable, like the amount of input DNA.


    1. Add the following components directly to the End Prep reaction mixture and mix well: 
      Blunt/TA Ligase Master Mix   15 μl 
      NEBNext Adaptor for Illumina*   2.5 μl 
      Ligation Enhancer   1 μl 
      Total volume   83.5 μl

      *The NEBNext adaptor is provided in NEBNext Singleplex (NEB #E7350) or Multiplex (NEB #E7335 #E7500) Oligos for Illumina.

    2. Mix by pipetting followed by a quick spin to collect all liquid from the sides of the tube.

    3. Incubate at 20°C for 15 minutes in a thermal cycler.

    4. Add 3 μl of USER™ enzyme to the ligation mixture from step 3.

    5. Mix well and incubate at 37°C for 15 minutes.