SequaGel MD Protocol

Gel Preparation:

  1. Preparing the Working Solutions:
    To cast a 0.8 to 1.0 mm thick gel (>40 cm vertical gel recommended) combine the following in an Erlenmeyer flask:
    • 50 ml SequaGel MD
    • 6 ml 10X TBE
    • 15 g Urea (optional)

    Fill to 100 ml with deionized water and mix thoroughly. Urea may assist the formation of more distinct bands during electrophoresis and reduces the formation of doublets in homoduplex controls.

  2. Casting the gel:
    Add the following to the solution, and swirl gently:
    • 40 μl TEMED
    • 400 μl freshly prepared 10% ammonium persulfate

    Treat one plate from the gel cassette with Glass Free to facilitate later disassembly of the cassette. Using standard acrylamide procedure, pour the gel solution into the cassette, insert the comb, and allow to polymerize at room temperature for a minimum of 60 minutes. Attach the gel cassette to the electrophoresis apparatus, and fill the upper and lower chambers with 0.6X TBE.

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