- Weigh and mix agarose powder with 1× TAE to desired agarose m/V percentage in a flask or bottle. An flask for agarose is usually by the gel station. Make enough volume for however many gels at their respective concentrations.
Alternatively, obtain previously prepared solid agarose gel in bottle/flask. - With a loosened cap, microwave agarose suspension or solid until completely clear and homogenous liquid (≈1 min per 100 mL on high). Watch as it boils to stop the microwave when it begins to bubble up. Swirl, handling with a silicone mit. Continue microwaving in small increments if necessary.
- If desired, dilute molten agarose with TAE to desired percentage, swirling to mix. Make enough for the desired casts and the level you wish to fill. See below.
For example, you can keep a bottle of 2% agarose and melt it as needed, diluting in a second flask to the desired 0.8–2% concentration. Pouring in a second container and dilution also cools it, so the poured gel solidifies faster. - If pre-staining the gel, add 10,000× DNA dye (GelGreen or GelRed) to molten agarose and swirl until color is uniform.
- Pick gel casting trays appropriate for your use. The short tray holds 40 mL when filled to the top of comb wells. The long tray holds 55 mL when filled to the top of the comb wells.
For gel purification purposes, you may want a thick gel (fully filled cast) to minimize the number of wells needed to hold the samples. For analytical purposes, you may need only a thin, half-filled gel. - Position trays in center of clamp and tighten clamp moderately. Place desired comb(s) in tray slots, oriented so the prongs are closer to the top of the gel.
- Pour molten agarose in tray to desired level while still hot, but not boiling to prevent tray/clamp warping. Use comb to pull bubbles away to bottom of gel.
- Allow the gel to solidify at room temperature or in the deli fridge 8–15min. Don't take combs out too early, or the wells will collapse.
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