Use the tables below corresponding to the AquaPor agarose of choice to determine the percentage gel to cast. (LE = general purpose; LM = low melting; ES = extra strength; HR = high resolution).
AquaPor LE
(all around performance) | | Gel % | Size Range (bp) | | 2 | 200-3,000 | | 1.75 | 250-4,000 | | 1.5 | 300-5,000 | | 1 | 400-12,000 | | 0.75 | 1,000-23,000 |
| AquaPor LM
(low melting) | | Gel % | Size Range (bp) | | 4 | <500 | | 3 | 200-800 | | 2 | 300-3,000 | | 1 | 500-20,000 | | 0.75 | 600-25,000 |
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AquaPor ES
(extra strength) | | Gel % | Size Range (bp) | | 1.5 | 300-5,000 | | 1 | 400-12,000 | | 0.75 | 800-15,000 | | 0.5 | 1,000-25,000 | | 0.3 | 5,000-50,000 |
| AquaPor HR
(high resolution) | | Gel % | Size Range (bp) | | 4.5 | 50-200 | | 4 | 75-300 | | 3 | 100-700 | | 2.5 | 125-800 | | 2 | 150-1,000 |
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Preparing an Agarose Gel
Before preparing an agarose gel it is first important to select a quality comb and the correct buffer. Use a thin comb (less than 1 mm) with wide teeth for the sharpest, best resolved bands.
Buffer selection depends on the condition and/or use of DNA. We recommend the following guidelines:
- Use 1X TBE for optimal resolution of DNA < 12 kb when the DNA will not be recovered.
- Use 1X TAE for the best separation of DNA from 12 kb to 50 kb, or for DNA < 12 kb if the DNA will be recovered from the gel.
- Use 1X Tris-Acetate (TAE without EDTA) if the DNA will be used for in-gel enzymatic processing.
Casting a Gel- Add buffer (at room temperature) to a flask that is 2.5 - 4 times the volume of gel solution. Add a teflon-coated stir bar.
- Add AquaPor powder while stirring vigorously so the agarose is dispersed uniformly. Stir for 2 minutes to hydrate the agarose.
- Tare the flask and solution.
- Place in a microwave oven and heat at 100% power using 20-60 second intervals. Swirl gently between intervals to resuspend the agarose.
- Continue the cycle of heating and swirling until the agarose is completely dissolved (no visible particles are present).
- Add distilled water to return the solution to its initial weight and mix.
- Cool the solution to 50-60°C before pouring the gel. Pour the solution into the mold so as to dispense the entire amount in 30-60 seconds, without generating bubbles.
- After casting, chill the gel for 30 minutes prior to comb removal when using AquaPor LM, HR, and low (<1%) concentrations of AquaPor LE and ES. This will complete gelation, increase gel strength, and enhance DNA resolution.
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