PCR Analysis of Tail DNA: Difference between revisions
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==Protocol== | ==Protocol== | ||
#Use the following Volumes per Reaction: | |||
Use the following Volumes per Reaction: | #Buffer: 4 uL of 5X Go-Taq buffer ("Molecular Biology Stuff" box in freezer) | ||
#Forward Primer: .4ul | |||
Buffer: 4 uL of 5X Go-Taq buffer ("Molecular Biology Stuff" box in freezer) | #Reverse Primer: .4ul | ||
#dNTPs: .4uL of 2 mM ("Molecular Biology Stuff" box in freezer) | |||
Forward Primer: .4ul | #Sterile water: 13.6 uL | ||
#Polymerase Go-Taq: 1 uL (6th floor in Genotype Yellow Box in freezer) | |||
Reverse Primer: .4ul | #Template: 1 uL | ||
dNTPs: .4uL of 2 mM ("Molecular Biology Stuff" box in freezer) | |||
Sterile water: 13.6 uL | |||
Polymerase Go-Taq: 1 uL (6th floor in Genotype Yellow Box in freezer) | |||
Template: 1 uL | |||
Run PCR Program (approx 2 hours). | Run PCR Program (approx 2 hours). | ||
Use Cycler 1 on 6th Floor | Use Cycler 1 on 6th Floor | ||
*Login: Sergey, Just press enter to Login | |||
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 | |||
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 | 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 | see [[Preparing an Agarose Gel]] for details on preparing a DNA gel | ||