View source for Preparation of Tail Samples (for Genotyping)

==PBND Solution: Tail Lysis Buffer==
* 50mM KCl                           (Hint: KCl is very heavy, you'll only need a tiny bit)
* 10mM Tris HCl (pH~8.3)
* 0.1 mg/mL MgCl2, 6H20
* 0.1 mg/mL Gelatin
* 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.

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


[[Category: Mouse Work]]
[[Category: Molecular Biology]]
[[Category: DNA]]
[[Category: Genotyping]]