Changes

* 50mM KCl (Hint: KCl is very heavy, you'll only need a tiny bit)

* 0.45% (NP-40) Jgepal (Hint: Use cut tips, as NP-40 is very viscous)* 0.45% Tween 20 (Hint: Use cut tips, as Tween 20 is very viscous) PBND can be made in liter volume, aliquoted into 50ml falcon tubes, frozen, and used as needed For 250ml PBND:* 0.930g 50mM KCl* 0.300g 10mM Tris HCl (pH~8.3)* 0.025g 0.1 mg/mL MgCl2, 6H20* 0.025g 0.1 mg/mL Gelatin* 1.125ml 0.45% (NP-40) Jgepal * 1.125ml 0.45% Tween 20  # Start by adding 150ml ddH2O to a beaker on a stir plate (with magnetic stir bar).# Add reagents. Mix well.# Add 37% HCl drop-wise to adjust pH to 8.3.# Add ddH20 to 250ml.

# Combine 100-200uL 100uL of PBND solution with 2ul of Proteinase K and mouse tail in an eppindorf eppendorf tube or in a well of a 96 well PCR plageplate (Proteinase K stock = 10mg/mL in ddH2O)# Incubate at 55 degrees (16 hours - O/N)# Incubate at 85 degrees for 45 60 min # Run PCRHold at 4 degrees