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  • NEBNext Quick Ligation Module Protocol (E6060)


    1. Mix the following components in a sterile microfuge tube: 
      End Repaired, Blunt DNA:   50 μl  
      NEBNext Quick Ligation Reaction Buffer (5X):   40 μl 
      *P1 Adaptor (50 μM):   variable
      *P2 Adaptor (50 μM):   variable
      T4 DNA Ligase:   10 μl 
      Sterile H2O:   variable 
      Total volume:   200 μl

      *Adaptors are not included, use adaptors appropriate to specific application. Adjust adaptor cocentration to obtain a final adaptor to DNA ratio of 30:1.
    2. Incubate in a thermal cycler for 15 minutes at 20°C.
    3. Purify DNA sample on one column, elute in 50 μl of water or EB buffer.
    4. Size select library fragments in the 200-230 bp range. Size selection can be performed using a number of methods including E-gel size select gels or standard 2% agarose gels.