2XFast Ligase Buffer

2X Quick Ligation Reaction Buffer: for 4 ml buffer recipe

132 mM Tris-HCL----- 528 ul autoclaved 1 M Tris-HCL (PH7.6)

20 mM MgCl2 ----- 80 ul autoclaved 1 M MgCL₂

2 mM dithiothreitol ----- 8 ul 1 M DTT (weight 0.154g DTT, dissolve with 1 ml autoclaved water, suggest that immediately store in the refrigerator after making this stock solution)

2 mM ATP ----- 800ul 10 mM ATP

15% Polyethylene glycol (PEG 6000) ----- 1.2 ml autoclaved 50% PEG 6000; 

pH 7.6 @25°C                                    ----- at last, add 1384 ul autoclaved water to make the final 4 ml buffer.