6X Crystal Violet Loading Dye 30% glycerol
20 mM EDTA
100 μg/mL crystal violet

Follow the instructions below to prepare a 0.8% agarose gel. The recipe will make one agarose gel with a volume of 50 ml.

  1. Mix 0.4 g of general purpose agarose and 50 ml 1X TAE buffer in a clean glass flask.
  2. Place flask in the microwave and heat until just boiling. Swirl to dissolve agarose and continue to heat in this fashion for 3 minutes to destroy nucleases.
  3. Remove from the microwave and allow to cool for 3 minutes.
  4. Add 20-40 μl of the 2 mg/ml Crystal Violet solution to the agarose and swirl to mix. The agarose should be light to medium purple in color.
  5. Rinse the gel box and comb with autoclaved water or TE buffer. Note: Use a comb that will hold all of the PCR amplification (48 μl) in one well, if possible.
  6. Pour the gel and set the comb in the gel.
  7. When the gel has solidified, cover the gel with 1X TAE buffer. You do not have to add crystal violet to the running buffer. Proceed to the next section to prepare your sample.