. Author manuscript; available in PMC: 2020 Apr 15.

Published in final edited form as: Methods. 2019 Feb 22;159-160:29–34. doi: 10.1016/j.ymeth.2019.02.019

Table 1.

Buffers and reagents used in this study.

Name	Components
Lysis buffer	50 mM Tris-HCl [pH 7.9], 1 mM EDTA, 10 μM ZnCl₂, 10% v/v glycerol, 1% v/v DMSO, 10 mM DTT and 1 × protease inhibitors
Ni buffer	20 mM Tris-HCl [pH 7.9], 150 mM KCl, 10 μM ZnCl₂, 10% v/v glycerol, 10 mM DTT and 1 × protease inhibitors
Ni wash buffer	20 mM Tris-HCl [pH 7.9], 150 mM KC1, 7 mM imidazole, 10 μM ZnCl₂, 10 mM DTT and 1 × protease inhibitors
MonoQ buffer	20 mM Tris-acetate [pH 7.9], 0.5 mM EDTA, 10 μM ZnCl₂, 10% v/v glycerol and 10 mM DTT
Elongation buffer	20 mM Tris-HCl [pH 8.0], 40 mM KC1, 5 mM DTT and 5 mM MgCl₂
Quench-loading buffer	90% (v/v) formamide, 50 mM EDTA, 0.05% (w/v) xylene cyanol and 0.05% (w/v) bromophenol blue
10X TBE buffer	1 M Tris base, 1 M Boric acid and 20 mM EDTA
TBE-Urea/PAGE	1 × TBE, 7 M urea and 8–16 % (v/v) bis-aerylamide 19:1
Exchange buffer	25 mM Tris [pH 7.5], 20 mM NaCl, 5 mM DTT, 1 μM Zn(OAc)₂ and 100 μM EDTA
Crystallization buffer	390 mM Ammonium phosphate [pH 6.0], 5 mM DTT, 5 mM Dioxane and 9–13 % (w/v) PEG 6,000
Cryo buffer	100 mM MES [pH 6.0], 350 mM NaCl, 5mM DTT, 5mM Dioxane, 16 % PEG 6,000 and 17 % PEG400