View source for PCR Analysis of Tail DNA

see [[Genotyping Details]] for strain specific details

==Materials==
# Dream Taq Green master mix
# Specific Primers
# DNA
# ddH2O

==Protocol==
Use the following Volumes per 50ul Reaction:

#10X GoTaq Buffer: 5uL ("Molecular Biology Stuff" box in freezer) 
#Primer Mix: 5ul
#dNTPs: 0.5uL of 10 mM ("Molecular Biology Stuff" box in freezer)
#Sterile water: 29ul 
#Polymerase Go-Taq: 0.125uL ("Molecular Biology Stuff"  box in freezer)
#Template: 1 uL

Master Mix (Per 5mL -- Make 1mL Aliquots)
#10X GoTaq Buffer: 625uL ("Molecular Biology Stuff" box in freezer) 
#Primer Mix: 625ul
#dNTPs: 62.5uL of 10 mM ("Molecular Biology Stuff" box in freezer)
#Sterile water: 3625ul 
#Polymerase Go-Taq: 15.625ul ("Molecular Biology Stuff"  box in freezer)
*Add Template Individually 

Run PCR Program (approx 2 hours).
Use Cycler 1 on 6th Floor
*Login: Sergey, Just press enter to Login
*Under Genotype folder, pick Ingles program for Ingles genotyping
*Under Genotype folder, pick regpcr program for PLT genotyping

Make sure to press enter 2x once to confirm Tubes and second time to start PCR

see [[Preparing an Agarose Gel]] for details on preparing a DNA gel

[[Category: Genotyping]]
[[Category: Mouse Work]]