NA Agarose Gel Electrophoresis – How to Run a DNA Agarose Gel

50x TAE
Tris 121g
EDTA 50 mL 500mm (EDTA pH to pH 8.0 with NaOH first before adding)
Acetic acid 28.55mL
Make up to 500ml

Make 1x TAE from 50x to water

NA Agarose Gel Electrophoresis – How to Run a DNA Agarose Gel

Pouring and Running agarose gel

  • disposable gloves when handling agarose gels, such as the highly toxic ethidium bromide
  • dispose of all waste ethidium bromide into a plastic bag (including pipette tips are touching the gel when you load a sample, the tape is to create a gel, etc.)
  • it may be necessary to clean the gel apparatus (combs and plastic holders) with water before use
  • for the small gel, using 25 ml of 1x TAE
  • for a large gel, using 50 ml of 1x TAE
  • generally, 1% agarose (0.25g in 25 mL) will work for most of the DNA fragment
  • for a larger fragment, a lower percentage of the agarose can be used, and for the smaller fragments, a higher percentage of the agarose can be used (0.4% to 2%)
  • using a thermos designated to agarose gel to make them
  • pour in 1x TAE, and then add the agarose
  • the microwave for 30 seconds to gel a little, and 60 seconds for a large gel
  • wait for cold gel (to speed up the cooling, it is possible to run a pumpkin under cold tap water)
  • while the cooling gel, place tape on both ends of the plastic holder to gel and put the comb in
  • when the solution has been cooled to about the temperature of your hand, add ethidium bromide (2uL for small gel, 4UL for large gel)
  • rinse the flask with hot water several times
  • when the gel is ready to run, remove the tape, put gel into the gel box, and then remove the comb (remember to rinse)
  • when running the gel, is set to about 100V (unplug the wire in the other apparatus to prevent overheating of the solution in other equipment); when extracting the gel, regulates the voltage to 80V
  • after the gel is ready for viewing, turn off the voltage, place it into a plastic tray, and take it to the camera

The ethidium bromide was prepared by adding 50uL of ethidium bromide is concentrated to 450uL of 1x TAE