Protocol for Oligonucleotide Adenylation (NEB #E2610)

Protocol

1. Set up the following reaction in a sterile microfuge tube:

   Components	Volume
   Phosphorylated DNA Oligonucleotide	100 pmol (5 pmol/µl)
   10X 5´ DNA Adenylation Reaction Buffer	2 µl
   1 mM ATP	2 µl
   Mth RNA Ligase	2 µl (100 pmol)
   Nuclease-free Water	to 20 µl

2. Incubate at 65°C for 1 hour.
3. Inactivate the enzyme by incubation at 85°C for 5 minutes.