NEBNext End Prep (E7442)
Protocol
Starting Material: 5 ng–1 μg fragmented DNA- Mix the following components in a sterile, nuclease-free tube:
End Prep Enzyme Mix 3.0 μl
End Repair Reaction Buffer (10X) 6.5 μl
Fragmented DNA 55.5 μl
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Total volume 65 μl - Mix by pipetting, followed by a quick spin to collect all liquid from the sides of the tube.
- Place in a thermocycler, with the heated lid on, and run the following program:
30 minutes @ 20°C
30 minutes @ 65°C
Hold at 4°C - Proceed directly to NEBNext Ultra Ligation Module (NEB #E7445).
