Do annealed double-stranded oligos need to be modified prior to ligation into the appropriate restriction site of my cloning vector?
Our chemical synthesis results in a hydroxyl group on both the 3' and 5' terminus of the oligo. DNA ligase requires a 5' phosphate and a 3' hydroxyl group for activity, so a 5' phosphate will need to be added if the intent is to ligate the oligo. This can be requested upon order using the modification code /5Phos/ in the sequence of your oligo. Alternatively, this modification can be added enzymatically through a Polynulcleotide Kinase (PNK) reaction.