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