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  • Protocol for RNA (E7445)

    Symbols
     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.

    Protocol

    Perform Adaptor Ligation
    1. Add the following components directly to the End Prep reaction mixture (65 μl) and mix well:
      Blunt/TA Ligase Master Mix   15 μl
      NEBNext Adaptor (15 μM)*   1 μl
      Nuclease-free Water   2.5 μl
      ----------------------------------------------------
      Total volume   83.5 μl
      *The adaptor is provided in NEBNext Singleplex (NEB #E7350) or NEBNext 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 15 minutes at 20°C.
      Note: USER step is performed during the PCR reaction.
    4. Proceed to desired size selection method.