Protocol for RNA (E7445)

 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.


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.