Annealing oligo dNTP linkers
- Mix the following:
- 45 μl of each linker strand dissolved in H2O at 10 pmol/μl
- 10 μl of 10x Annealing buffer
- 250 mM Tris-HCl, pH 8.0
- 100 mM MgCl2
- Boil a beaker of water. Turn off the flame, and put the tube in the water.
- Allow to cool to room temperature.
Use about 0.5 μl for a 10 μl ligation.
I have had problems sometimes with this mixture inhibiting ligation. I that case, ethanol precipitation (1/10 volume NaAc, 2.5 volume EtOH) followed by resuspension in TE solved the problem.