Difference between revisions of "PCR Analysis of Tail DNA"
From Bridges Lab Protocols
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==Protocol== | ==Protocol== | ||
| − | Use the following Volumes per Reaction: | + | Use the following Volumes per 50ul Reaction: |
| − | #Buffer: | + | #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) | |
| − | #dNTPs: 0. | + | #Sterile water: 29ul |
| − | #Sterile water: | + | #Polymerase Go-Taq: 0.125uL ("Molecular Biology Stuff" box in freezer) |
| − | #Polymerase Go-Taq: | + | |
#Template: 1 uL | #Template: 1 uL | ||
| + | |||
| + | Master Mix | ||
| + | #10X GoTaq Buffer: 50uL ("Molecular Biology Stuff" box in freezer) | ||
| + | #Primer Mix: 50ul | ||
| + | #dNTPs: 5uL of 10 mM ("Molecular Biology Stuff" box in freezer) | ||
| + | #Sterile water: 290ul | ||
| + | #Polymerase Go-Taq: 1.25ul ("Molecular Biology Stuff" box in freezer) | ||
| + | *Add Template Individually | ||
Run PCR Program (approx 2 hours). | Run PCR Program (approx 2 hours). | ||
Revision as of 15:00, 25 July 2013
see Genotyping Details for strain specific details
Materials
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
- 10X GoTaq Buffer: 50uL ("Molecular Biology Stuff" box in freezer)
- Primer Mix: 50ul
- dNTPs: 5uL of 10 mM ("Molecular Biology Stuff" box in freezer)
- Sterile water: 290ul
- Polymerase Go-Taq: 1.25ul ("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
