dA-Tailing of cDNA Library (E6110)

Protocol

1. Mix the following components in a sterile 1.5 ml microcentrifuge tube:
   Purified, End Repaired cDNA: 42 μl
   10X NEBNext dA-Tailing Reaction Buffer: 5 μl
   Klenow Fragment (3´→5´ exo^–): 3 μl
   -----------------------------------------------------------------
   Total volume: 50 μl
2. Incubate in a heat block for 30 minutes at 37°C.
3. Purify dA-Tailed cDNA using a PCR column purification kit, purifying the sample on one column and eluting in 38 μl sterile water or elution buffer.