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Directions for gel electrophoresis lab prep: mdna [Enough for 10 gels/50 mL each-70 students/1 ladder lane per gels]


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Directions for gel electrophoresis lab prep: mDNA [Enough for 10 gels/50 mL each-70 students/1 ladder lane per gels]
Prepare 7 liters of 1% TBE buffer-do this in 2 or 3 L flasks:
1. Mix 50 mL of 20X TBE with distilled water UP TO 1000mL for 1% TBE.

Mix 1-2 minutes.
2. Set aside 1 liter for GEL preparation.
3. For the other 6 Liters of 1xTBE, add gel stain (blue) drops—24 drops per each 500 mL.
4. Refrigerate buffers.

Prep and pour agarose gels:


  1. DOUBLE tape the ends of all 20 gel boxes.



  2. Prepare agarose in 2-500 mL flasks:
    Add 5 grams agarose to 333 mL 1% TBE buffer [NON-stained/prepared above] in a clean 1L flask. Cover with aluminum foil and heat in a boiling water bath on a hot plate until the agarose powder dissolves—10-20 minutes. Solution gets clear as it heats. Swirl and observe bottom to insure that ALL agarose is completely dissolved.



  3. Monitor the temperature of the solution and allow it to cool to around 65 degrees Celsius before pouring into gel boxes. Add 20 drops of gel stain to each 500 mL sample.



  4. Pour 50 mL of agarose into each gel box and place a white plastic comb into each after pouring—into the comb slit nearest the END of the box/black line.
    This should fill 20 gel boxes.



  5. If agarose starts to solidify, stop pouring and reheat to get back to liquid state---don’t pour “lumpy” agarose.



  6. Store each gel/comb set up in a separate Ziploc bag and refrigerate.


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