Standard PCR (25 ul reactions):
- 13.4 ul H20
- 2.5 ul 10X PCR Buffer
- 1.5 ul MgCl2 (2.5 mM)
- 1.0 ul Forward Primer (10uM)
- 1.0 ul Reverse Primer (10uM)
- 0.5 ul dNTPs (2.5mM each)
- 0.1 ul Taq Polymerase (5 U/ul)
- 5 ul DNA Template
- Concentrations refer to the reagent stock solutions, not the reaction itself.
- Keep Taq in the freezer until you need it. Add it last.
- Spin the Taq briefly before using.
- Mix all the reagents, exluding the template, into a cocktail.
- Make enough cocktail for an extra reaction or two.
- Put 20 ul of cocktail in each tube (on ice).
- Add 5ul template to each tube, pipet up and down.
Department of Biology, PO Box 400328
University of Virginia, Charlottesville, VA 22904-4328
Email: drt3b@virginia.edu Phone:(434)982-5217
Site Design by Eyewall Design.com & Gilad Aharoni
