- Mix	1 μl annealed oligos
	1 μl 10× T4 DNA ligase buffer (NEB)
	7 μl water
	1 μl T4 PNK (polynucleotide kinase, NEB)
	(if using PNK buffer, the reaction needs to be supplemented with ATP)
- Incubate 30 min at 37°C for the oligo phosphorylation, then 10 min at 70°C to inactivate the enzyme.
- The phosphorylated oligos are now at 5 μM.
- Dilute an aliquot, e.g. 5 μl, 1:125 with water to a final conc. of 40 nM.