Classic SDS-PAGE
STOCK SOLUTIONS:
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a b c d e f
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ACRYLAMIDE 30g - - - - -
BIS 0,8g - - - - -
TRIS - 18,17g 6,06g 12,0g - -
6N HCl - p/ pH8,8 p/pH6,8 - - -
GLICINA - - - 57,6g - -
10% SDS - - - - 2,6ml -
DTT - - - - 96,3mg -
PERSULFATE - - - - - 0,5g
(c) - - - - 1,25ml -
GLYCEROL 30g - - - 1,26g -
0.5M EDTA (Na4) - - - - 10ul -
0.2M EGTA - - - - 25ul -
0.1% Bromophenol Blue - - - - 1,5ml -
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Water to 100 ml 100 ml 100 ml 1000ml 10 ml 5,0ml
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a=Bis-Acrylamide; b=Buffer b; c=Buffer c; d=Tris-Gly 4x; e=Sample Buffer;
e=10% (w/v) Ammonium Persulfate (AP).
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WORKING SOLUTIONS (for Bio-Rad small apparatus):
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STACKING GEL 10% RUNNING GEL 15% RUNNING GEL
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Water 3,10 ml 4,30 ml 2,63 ml
Solution a 500 ul 3,33 ml 5,00 ml
Buffer b - 2,50 ml 2,50 ml
Buffer c 1,25 ml - -
10% SDS 49,3 ul 100 ul 100 ul
10% Persulfato 15,0 ul 45,0 ul 45,0 ul
TEMED 10,0 ul 7,5 ul 7,5 ul
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FINAL VOLUME 4,924 ml 10,283 ml 10,283 ml
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