Turning Oligos into Linkers

1. dilute oligos to 1mg/ml in dH2O
2. 2.5mg oligos (each) into 100ml 1x kinase buffer with 1mM ATP

oligo 1 2.5ml (1mg/ml)

oligo 2 –2.5ml (1mg/ml)

10x kinase buffer—10ml

10mM ATP—10ml

H2O—75ml

3. add 1ml T4 polymerase kinase (10unuits) and incubate 1 hour at 37 C
4. add 3ml 0.1M EDTA, 1ml 20x SSC, heat to 65 C (to kill kinase)
5. Incubate at 65C for 10-20 minutes. Turn off water bath until RT.

(alternatively, reaction mixture—100C on heat block –turn off –until 10C below Tm)