Team:Washington/Protocols/PAGE gel electrophoresis

From 2012.igem.org

(Difference between revisions)
(→General Procedure)
(→Equipment Necessary)
 
(5 intermediate revisions not shown)
Line 9: Line 9:
#Remove water, add staining agent (GelCode Blue), leave on rocker overnight
#Remove water, add staining agent (GelCode Blue), leave on rocker overnight
#Remove GelCode Blue, add DI water. Refresh DI water every hour until gel bands are visible
#Remove GelCode Blue, add DI water. Refresh DI water every hour until gel bands are visible
 +
 +
 +
== SDS PAGE Gel Buffer Information ==
 +
 +
4X Loading Buffer
 +
 +
#2.0 ml 1M Tris-HCl pH 6.8
 +
#0.8 g SDS
 +
#4.0 ml 100% glycerol
 +
#0.4 ml 14.7 M β-mercaptoethanol
 +
#1.0 ml 0.5 M EDTA
 +
#8 mg bromophenol Blue
 +
 +
 +
 +
10X Running Buffer
 +
 +
#288 g glycine
 +
#60.4 g Tris base
 +
#20 g SDS
 +
#1.8 L H2O
 +
 +
 +
 +
#Take protein sample, add Loading Buffer. Boil at 95 degrees Celsius for ten minutes
 +
#Remove premade gel cartridge from package, remove comb, rinse wells with DI water
 +
#Insert cartridge into gel box, add running buffer to top and bottom wells
 +
#Add samples into separate wells, along with a protein ladder.
 +
#Run gel at 200V for around one hour
 +
 +
== Equipment Necessary ==
 +
 +
#Power Supply
 +
#SDS-PAGE Gel Box
 +
#SDS-PAGE Gel Cartridge
 +
 +
 +
'''← [[Team:Washington/Protocols|Back to Protocols]]'''

Latest revision as of 02:13, 4 October 2012

Contents

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

Equipment Necessary

  1. Power Supply
  2. SDS-PAGE Gel Box
  3. SDS-PAGE Gel Cartridge


← Back to Protocols

Retrieved from "http://2012.igem.org/Team:Washington/Protocols/PAGE_gel_electrophoresis"