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Top Questions

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.
Tags:
  • ligate
  • hydroxyl
  • ligation
  • 5' phosphate
  • ligase
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