Tail Digestion for DNA in 96-well plate Protocol
Add 200 µl of 0.05M NaOH1 (Sodium Hydroxide) to wells in 96-well PCR plate3.
Add tail snips to wells.
Incubate in a thermocycler at 98oC for 20 minutes.
Remove from heat. Add 20 µl of 1M Tris-HCl2 (pH 7.5)3.
Spin plates at 2000 RPMs for 7 minutes to settle debris.
Ready to use in PCR.
10.05M NaOH
mix 2 grams NaOH (F.W. 40.00) in 1 liter 18 MOhm H2O (not autoclaved)
21M Tris-HCl
mix 78.8 grams Tris-HCl into a final volume of 500 ml 18 MOhm water, to pH 7.5 using NaOH
3I usually load plates using a multi-channel pipetter, whenever practical.
If any questions, contact Tom Sproule @ tom.sproule@jax.org.
