NEBNext Quick Ligation Module Protocol (E6060)

Protocol

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 
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   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.