logo
AAT Bioquest

What are the steps of the gel electrophoresis process?

Posted February 9, 2022


Answer

The gel electrophoresis consists of 8 steps:

  1. Preparing the samples for running – The DNA is isolated and blue dye is added to the solution to make it easy to visualize the movement of the sample through the gel.
  2. Preparing an agarose TAE gel solution – TAE buffer solution is necessary for generating an electric field during electrophoresis. A weight-to-volume concentration of agarose in the TAE buffer is used to prepare the solution, which is then heated to dissolve the agarose.
  3. Casting the gel – The agarose TAE solution is poured into a casting tray and allowed to cool and solidify, which creates a gel slab with a row of wells at the top.
  4. Setting up the electrophoresis chamber – This is done by placing the solid gel into a chamber filled with the TAE buffer. The gel is placed so that it is in closest proximity to the negative electrode of the chamber.
  5. Loading the gel – The gel chamber wells are loaded with the DNA sample along with a DNA ladder, which is used as a reference for sizes.
  6. Initiating electrophoresis – The positive and negative points are connected to the power supply and chamber and the power is switched on to generate an electric field. The negatively charged DNA fragments will move away from the negative electrode and towards the positive electrode.
  7. Stopping electrophoresis: – The power supply off is turned off when the blue-stained DNA samples have migrated far enough through the gel. The gel is removed. 
  8. Visualizing the DNA – Once the fragments have been separated, the gel is placed in an ethidium bromide solution. The ethidium bromide stained gel is then exposed to UV light and a picture is taken. DNA bands appear in each lane corresponding to a chamber well. The DNA ladder is also visible, which helps in determining the length of the DNA bands.
Additional resources

Sample preparation of gel electrophoretically separated protein binding partners for analysis by mass spectrometry

Gelite™ Safe DNA Gel Stain *10,000X DMSO Solution*