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?
-tedijp-
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.
QUOTE (tedijp @ Sep 4 2006, 02:49 PM)
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?


-tertu-