Protocol Online logo
Top : Forum Archives: : Molecular Biology

Ammonium Sulfate PCR buffer - Ammonium Sulfate PCR buffer (Feb/24/2003 )

I am looking for a recipe for ammounium sulfate PCR buffer. A buffer that is compatible with Real-time PCR would be preferred. Thank you to all who respond.

-dnanerd-

An ammonium sulfate PCR buffer formula from New England Biolabs:
10x NEB ThermoPol Reaction buffer: 0.2 mM Tris-HCl, pH 8.8 0.05 @ 25C, 0.1 M KCl, 0.1 M (NH4)2SO4, 20 mM MgSO4, 1% (v/v) Triton X-100.

100 mM MgSO4 solution is provided by NEB in case a higher level of magnesium is required.

1X TaqMan Buffer A (Formulation available from PE, but basically PCR buffer plus
ROX to 60nM at 1X Concentration):
7.5 mM MgCl2
200 M each dATP, dCTP, dGTP
400 M dUTP
0.05 U/uL AmpliTaq Gold
0.01 U/uL Amperase UNG

Combine aspects of the two buffers (higher magnesium for real-time PCR),

-tfitzwater-