This is a cached page for the URL (http://www.research.umbc.edu/~jwolf/m6.htm#agarose). To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
Agarose Gel Electrophoresis M.6: AGAROSE GEL ELECTROPHORESIS                                                  Go Home
Materials:
1. To prepare 100 ml of a 0.7% agarose solution, measure 0.7 g agarose into a glass beaker or flask and add 100 ml 1X TBE or TAE.
2. Microwave or stir on a hot plate until agarose is dissolved and solution is clear.
3. Allow solution to cool to about 55C before pouring. (Ethidium bromide can be added at this point to a concentration of 0.5 µg/ml)
4. Prepare gel tray by sealing ends with tape or other custom-made dam.
5. Place comb in gel tray about 1 inch from one end of the tray and position the comb vertically such that the teeth are about 1-2 mm above the surface of the tray.
6. Pour 50C gel solution into tray to a depth of about 5 mm. Allow gel to solidify about 20 minutes at room temperature.
7. To run, gently remove the comb, place tray in electrophoresis chamber, and cover (just until wells are submerged) with electrophoresis buffer (the same buffer used to prepare the agarose)
8. Excess agarose can be stored at room temperature and remelted in a microwave.
9. To prepare samples for electrophoresis, add 1 µl of 6x gel loading dye for every 5 µl of DNA solution. Mix well. Load 5-12 µl of DNA per well (for minigel).
10. Electrophorese at 50-150 volts until dye markers have migrated an appropriate distance, depending on the size of DNA to be visualized.
11. If the gel was not stained with ethidium during the run, stain the gel in 0.5 µg/ml ethidium bromide until the DNA has taken up the dye and is visible under short wave UV light, if the DNA will not be used further, or with a hand-held long-wave light if the DNA is to be cloned.

 
50x TAE
242 g Tris base
57.1 g glacial acetic acid
100 ml 0.5 M EDTA
10x TBE
108 g Tris base
55 g boric acid
40 ml 0.5 M EDTA, pH=8
distilled water to 1 liter
6x gel loading buffer
0.25% Bromophenol blue
0.25%Xylene cyanol FF
15% Ficoll Type 4000
120 mM EDTA