Component	Final concentration	Stock	Amount
Tris–HCl (pH 7.5)	500 mM	1 M	2.5 ml
MgCl2	100 mM	1 M	500 μl
Dithiothreitol	100 mM	1 M	500 μl
ATP	10 mM	100 mM	500 μl
dATP	4 mM	100 mM	200 μl
dCTP	4 mM	100 mM	200 μl
dGTP	4 mM	100 mM	200 μl
dTTP	4 mM	100 mM	200 μl

Add water to 5 ml

Note: This buffer can be purchased from New England Biolabs as part of the NEBNext Master Mix 3 (cat# E6060S/L) or as an individual buffer (cat# E6062A/AA)