PCR Analysis of Tail DNA: Difference between revisions
Jump to navigation
Jump to search
No edit summary |
mNo edit summary |
||
| Line 4: | Line 4: | ||
==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 | ||
Revision as of 17:59, 5 June 2009
see Genotyping Details for strain specific details
Materials
Protocol
- Use the following Volumes per Reaction:
- Buffer: 4 uL of 5X Go-Taq buffer ("Molecular Biology Stuff" box in freezer)
- Forward Primer: .4ul
- Reverse Primer: .4ul
- 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). 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