SDS-PAGE

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(Solutions for various polyacrylamide gels (SDS-PAGE))
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===Solutions for 10 ml (MINIgel) of various polyacrylamide gels (SDS-PAGE)===
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===Running (lower) gel: 10 ml (2 MINIgels)===
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  Chemical      8%    10%    12%
  Chemical      8%    10%    12%
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  Water        4.74  4.0    3.3  ml
  Water        4.74  4.0    3.3  ml
  30% AA(*)    2.66  3.3    4.0  ml
  30% AA(*)    2.66  3.3    4.0  ml
  LOWER buffer  2.5    2.5    2.5  ml
  LOWER buffer  2.5    2.5    2.5  ml
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20% [[SDS]]      50    50      50  ul
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10% APS      200    200    200  ul
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TEMED        8.2    8.2    8.2  ul
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===Stacking (upper) gel: 4 ml (2 MINIgels)===
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Chemical      4%   
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Water        2.65 ml
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30% AA(*)    0.67 ml
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UPPER buffer  0.65 ml
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20% [[SDS]]      20 ul
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10% APS      125 ul
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TEMED        6 ul
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===Buffers===
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LOWER buffer: 1M Tris base, pH 8.8
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UPPER buffer: 1M Tris base, pH 6.8
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===Abbreviations===
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AA = acrylamide
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[[SDS]] = sodium dodecylsulfate
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APS = ammonium persulfate
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==Notes==
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[http://www.ruf.rice.edu/~bioslabs/studies/sds-page/sdsgoofs.html SDS-PAGE "Hall of Shame"] - examples of common failures with debugging tips.
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[[Category:Material]] [[Category:PAGE]]

Revision as of 10:48, 8 April 2009

Contents

Running (lower) gel: 10 ml (2 MINIgels)

Chemical      8%     10%     12%

Water         4.74   4.0     3.3  ml
30% AA(*)     2.66   3.3     4.0  ml
LOWER buffer  2.5    2.5     2.5  ml
20% SDS       50     50      50   ul
10% APS       200    200     200  ul
TEMED         8.2    8.2     8.2  ul

Stacking (upper) gel: 4 ml (2 MINIgels)

Chemical      4%     

Water         2.65 ml
30% AA(*)     0.67 ml
UPPER buffer  0.65 ml
20% SDS       20 ul
10% APS       125 ul
TEMED         6 ul

Buffers

LOWER buffer: 1M Tris base, pH 8.8
UPPER buffer: 1M Tris base, pH 6.8

Abbreviations

AA = acrylamide
SDS = sodium dodecylsulfate
APS = ammonium persulfate

Notes

SDS-PAGE "Hall of Shame" - examples of common failures with debugging tips.

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