Team:Washington/Protocols/PAGE gel electrophoresis

From 2012.igem.org

(Difference between revisions)
(→SDS PAGE Gel Buffer Information)
(→SDS PAGE Gel Buffer Information)
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4X Loading Buffer
4X Loading Buffer
-
2.0 ml 1M Tris-HCl pH 6.8
+
#2.0 ml 1M Tris-HCl pH 6.8
-
 
+
#0.8 g SDS
-
0.8 g SDS
+
#4.0 ml 100% glycerol
-
 
+
#0.4 ml 14.7 M β-mercaptoethanol
-
4.0 ml 100% glycerol
+
#1.0 ml 0.5 M EDTA
-
 
+
#8 mg bromophenol Blue  
-
0.4 ml 14.7 M β-mercaptoethanol
+
-
 
+
-
1.0 ml 0.5 M EDTA
+
-
 
+
-
8 mg bromophenol Blue  
+
10X Running Buffer
10X Running Buffer
-
288 g glycine
+
#288 g glycine
-
 
+
#60.4 g Tris base
-
60.4 g Tris base
+
#20 g SDS
 +
#1.8 L H2O
-
20 g SDS
 
-
1.8 L H2O
 
#Take protein sample, add Loading Buffer. Boil at 95 degrees Celsius for ten minutes
#Take protein sample, add Loading Buffer. Boil at 95 degrees Celsius for ten minutes

Revision as of 00:26, 3 October 2012

SDS PAGE Gel Electrophoresis

General Procedure

  1. Place gel cartridge in gel box with loading buffer
  2. Load samples
  3. Run the gel
  4. Place gel in DI water for one hour
  5. Remove water, add staining agent (GelCode Blue), leave on rocker overnight
  6. Remove GelCode Blue, add DI water. Refresh DI water every hour until gel bands are visible


SDS PAGE Gel Buffer Information

4X Loading Buffer

  1. 2.0 ml 1M Tris-HCl pH 6.8
  2. 0.8 g SDS
  3. 4.0 ml 100% glycerol
  4. 0.4 ml 14.7 M β-mercaptoethanol
  5. 1.0 ml 0.5 M EDTA
  6. 8 mg bromophenol Blue

10X Running Buffer

  1. 288 g glycine
  2. 60.4 g Tris base
  3. 20 g SDS
  4. 1.8 L H2O


  1. Take protein sample, add Loading Buffer. Boil at 95 degrees Celsius for ten minutes
  2. Remove premade gel cartridge from package, remove comb, rinse wells with DI water
  3. Insert cartridge into gel box, add running buffer to top and bottom wells
  4. Add samples into separate wells, along with a protein ladder.
  5. Run gel at 200V for around one hour
Retrieved from "http://2012.igem.org/Team:Washington/Protocols/PAGE_gel_electrophoresis"