FAQ: How much Mg2+ [magnesium(II)] should be included in a typical reaction?

NEB ThermoPol® Reaction Buffer contains 2mM Mg2SO4 (magnesium sulfate), which is suitable for most reactions carried out with TtAgo. Higher amounts of Mg2+ can improve TtAgo performance on some substrates, but may also lead to more aberrant, off-target activity. A similar trend of slightly higher overall performance accompanied by off-target activity is observed when using TtAgo with Mn2+ as with Mg2+, however Mn2+ has lower range (0.1 to 2mM) of working concentrations compared to Mg2+ (0.5 to 5mM). For more information, please refer to GUIDElines for optimization of Tth Argonaute (TtAgo) reactions.