The reaction buffer for Thermo Scientific Phire Hot Start II DNA Polymerase is available in three formats: non-colored version with and without detergents (F-524L and F-525L, respectively) or Green version (F-527L) containing a density reagent and two tracking dyes for direct loading of PCR products on gel. The colored buffer does not interfere with enzyme performance and is compatible with downstream applications such as DNA sequencing, ligation and restriction digestion.
The concentration of Mg2+ in the reaction buffers is optimized to work well for most amplicons. The buffer contains 1.5 mM MgCl2 at final reaction concentration. Separate tubes of MgCl2 and DMSO are provided for further optimization.