PCR Analysis of Tail DNA

see Genotyping Details for strain specific details

Protocol

Use the following Volumes per 50ul Reaction:

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

Master Mix (Per 5mL -- Make 1mL Aliquots)

1. 10X GoTaq Buffer: 625uL ("Molecular Biology Stuff" box in freezer)
2. Primer Mix: 625ul
3. dNTPs: 62.5uL of 10 mM ("Molecular Biology Stuff" box in freezer)
4. Sterile water: 3625ul
5. 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