End Repair of cDNA Library

For use with NEBNext® mRNA Library Prep Reagent Set for Illumina® (E6100) and NEBNext® mRNA Library Prep Master Mix Set for Illumina® (E6110) .


  1. Mix the following components in a sterile 1.5 ml microcentrifuge tube:
    Purified double-stranded cDNA: 50 μl
    Nuclease-free Water: 25 μl
    10X Phosphorylation Reaction Buffer: 10 μl
    Deoxynucleotide Solution Mix: 4 μl
    T4 DNA Polymerase: 5 μl
    E. coli DNA Polymerase I, Large (Klenow) Fragment: 1 μl
    T4 Polynucleotide Kinase: 5 μl 
    Total volume: 100 μl
  2. Incubate in a heat block for 30 minutes at 20°C.
  3. Purify end repaired DNA using a PCR column purification kit, purifying the sample on one column and eluting in 32 μl sterile water or elution buffer.