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Preparation of Tail Samples (for Genotyping)

6 bytes added, 15:20, 21 May 2018
changed g to ml for detergents
* 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)
PBDN 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
* 1.125g 125ml 0.45% (NP-40) Jgepal * 1.125g 125ml 0.45% Tween 20 
1. # Start by adding 150ml ddH2O to a beaker on a stir plate (with magnetic stir bar).2. # Add reagents. Mix well.3. # Add 37% HCl drop-wise to adjust pH to 8.3.4. # 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/mLin ddH2O)
# Incubate at 55 degrees (16 hours - O/N)
# Incubate at 85 degrees for 60 min
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Retrieved from "http://bridgeslab.sph.umich.edu/protocols/index.php/Special:MobileDiff/1372...1470"

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