5' phosphorylated primers - (Sep/04/2006 )
Hi! Can somebody tell me what the 5'phosphorylated primer is used for (except lambda-exonuclease treatment afterwards)? Is there a change in the PCR conditions when one primer is phosphorylated or the same protocol can be used?
You phosphorylate oligos if you are going to anneal them and use them for ligations as you need a 5' phosphate and a 3' OH.
For PCR, the 3' remains unphosphorylated so amplification takes place normally without further modifications.