Difference between revisions of "Preparation of Tail Samples (for Genotyping)"

From Bridges Lab Protocols
Jump to: navigation, search
m (added categories)
m (Protocol: Proteinase K)
Line 8: Line 8:
  
 
==Protocol==
 
==Protocol==
# Combine 100-200uL of PBND solution with 2ul of Proteinase K and mouse tail in an eppindorf tube or in a well of a 96 well PCR plage
+
# Combine 100-200uL of PBND solution with 2ul of Proteinase K and mouse tail in an eppindorf tube or in a well of a 96 well PCR plage (Proteinase K stock = 10mg/mL)
 
# Incubate at 55 degrees (O/N)
 
# Incubate at 55 degrees (O/N)
 
# Incubate at 85 degrees for 45 min  
 
# Incubate at 85 degrees for 45 min  

Revision as of 17:18, 26 June 2013

PBND Solution: Tail Lysis Buffer

  • 50mM KCl
  • 10mM Tris HCl (pH~8.3)
  • 0.1 mg/mL MgCl2, 6H20
  • 0.1 mg/mL Gelatin
  • 0.45% (NP-40) Jgepal
  • 0.45% Tween 20

Protocol

  1. Combine 100-200uL of PBND solution with 2ul of Proteinase K and mouse tail in an eppindorf tube or in a well of a 96 well PCR plage (Proteinase K stock = 10mg/mL)
  2. Incubate at 55 degrees (O/N)
  3. Incubate at 85 degrees for 45 min
  4. Run PCR