Difference between revisions of "SDS-PAGE"

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(Notes)
(SDS link)
Line 6: Line 6:
 
  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
  20% SDS      50    50      50  ul
+
  20% [[SDS]]       50    50      50  ul
 
  10% APS      200    200    200  ul
 
  10% APS      200    200    200  ul
 
  TEMED        8.2    8.2    8.2  ul
 
  TEMED        8.2    8.2    8.2  ul
Line 17: Line 17:
 
  30% AA(*)    0.67 ml
 
  30% AA(*)    0.67 ml
 
  UPPER buffer  0.65 ml
 
  UPPER buffer  0.65 ml
  20% SDS      20 ul
+
  20% [[SDS]]       20 ul
 
  10% APS      125 ul
 
  10% APS      125 ul
 
  TEMED        6 ul
 
  TEMED        6 ul
Line 29: Line 29:
  
 
  AA = acrylamide
 
  AA = acrylamide
  SDS = sodium dodecylsulfate
+
  [[SDS]] = sodium dodecylsulfate
 
  APS = ammonium persulfate
 
  APS = ammonium persulfate
  

Revision as of 07:48, 8 April 2009

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.