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RECIPES > TRICINE GEL RECIPE

> Preparation of Stock Solutions Required for Making Tricine SDS Gels

   

> Preparation of Tricine SDS Gels

   

> Preparation of the Stock Solutions Required for Running Tricine SDS Gels

    > Preparing to Run Tricine SDS Gels

Preparation of Tricine SDS Gels

  • Prepare Gel Cassettes
  • Prepare resolving gel (150 mLs; 30 gels) (for multicasters) (at lab bench)
    • In a 250 mL beaker add acrylamide stock and gel buffer
    • Add enough H20 to get 150mL
    • Degas the solution under a vacuum
    • Add polymerizing agents (AP and TEMED)
    • Pour solution between glass walls with syringe
    • Add a thin layer of 100% isopropanol with syringe. Pour off excess.
    • Allow to polymerize for 1-2 hour
 

10% Resolving gel - 150 mL

Acrylamide stock 33.4 mL
Gel buffer 50 mL
H2O

66.6 mL

10% AP 750 µL
TEMED 75 µL
  • Prepare stacking gel (60 mL) (at lab bench)
    • In a beaker add acrylamide stock and gel buffer
    • Add H2O
    • Degas the solution under a vacuum
    • Add polymerizing agents (AP and TEMED)
    • Pour solution between glass walls with syringe
    • Place combs on top.
    • Allow to polymerize 1-2 hour.
  4% Stacking gel - 60.4 mL
Acrylamide stock 5.4 mL
Gel buffer 15 mL
H2O 40 mL
10% AP 300 µL
TEMED 60 µL

 

Reference:

Hermann Schägger and Gebhard von Jagow, " Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa", Analyt. Biochem. 166, 368-379 (1987)