Purification after oligos phosphorylation? - (Dec/06/2006 )
Hi to all!
I want to insert one 55 bps to one 8 kbs vector.
I want to phosphorylate my oligos before annealing. Will I have to purify them before ligation to dephosphorylated and purified vector?
You can purify it by column or Qiagen kit, or You might simply heat 65C for 20 mins to inactive phosporylation before add to ligation.
while, I actually dislike the idea to phosphorylated such short insert by myself, synthezed oligos with 5' phosphorylated is more efficient!