Perform Adaptor Ligation (E7420)

Protocol

Dilute the NEBNext Adaptor for Illumina (15 μM) to 1.5 μM with a 10- fold dilution (1:9) with sterile water for immediate use.
  1. To the dA-Tailed cDNA (65 μl), add the following components:

    Blunt/TA Ligase Master Mix   15 μl
    Diluted NEBNext Adaptor   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. Incubate 15 minutes at 20°C.

    Note: USER™ step is performed during the PCR reaction.