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===Buffers List=== '''NOW USING 1.0 M NaCl AND 0.5% TRITON-X 100 FOR 1X HEGX, WASH BUFFER ONLY''' '''CLEAVAGE BUFFER WILL BE 0.8 M NaCl, AND 0.05% TRITON-X 100''' *1X HEGX 20 mM Tris HCl pH 7.5 1.0 M NaCl 1 mM EDTA 10% glycerol 0.5% Triton-X 100 *2X HEGX to make 1L 40 mM Tris HCl pH 7.5 20 mL of 2M 2.0 M NaCl 116.88 g 2 mM EDTA 0.58448 g or 4 mL of 500 mM 20% glycerol 200 mL 1.0% Triton-X 100 10 mL *2X Cleavage buffer (need to add DTT still): '''WILL BE 0.8 M NaCl, AND 0.05% TRITON-X 100 FINAL CONCENTRATION''' to make 50 mL 40 mM Tris HCl pH 7.5 1 mL of 2M 1.6 M NaCl 4.6752 g 2 mM EDTA 200 uL of 500 mM 0.1% Triton-X 100 50 uL 20% glycerol 10 mL *2xTn5 dialysis Bf: '''WITH TRIS-HCL INSTEAD''' 1L (H20 added to vol.) 100 mM Tris HCl, pH 7.5 50 mL 2M 200 mM NaCl 11.69 g NaCl 0.2 mM EDTA 400 uL 500 mM 2 mM DTT 2 mL 1M (309 mg) 0.05% Triton X-100 500 uL Triton X-100 20% Glycerol 252 g 100% Glycerol *Tn5 storage Bf: '''WITH TRIS-HCL INSTEAD''' 100 mL (+42 mL H20) 1L 2L 50 mM Tris HCl, pH 7.5 2.5 mL 2M 25 mL 2M 50 mL 2M 100 mM NaCl 2 mL 5M 5.844 g 11.69 g 0.1 mM EDTA 20 uL 500 mM 200 uL of .5M 400 uL of .5M 1 mM DTT 0.1 mL 1M 0.154 g 0.309 g 0.05% Triton X-100 500 uL 10% (25 uL 100%) 500 uL 100% 1000 uL 100% 50% Glycerol 63 g 100% Glycerol 500 mL 1000 mL
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