IGEM:PennState/2007/FreezeandSqueeze

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Freeze 'N Squeeze DNA Gel Extraction

1. Electrophorese the DNA sample in an ararose gel (TAE or TBE), then stain with an appropriate reagent, e.g., ethidium bromide or SYBR Green 1.

2. Using a clean razor blade, carefully excise the band of interest. Trim excess agarose from all six sides of the DNA band to maximize recovery and purity.

3. Chop the trimmed gel slice and place the pieces into the filter cup of the Quantum Prep Freeze 'N Squeeze DNA Gel Extraction Spin Column. Place the filter cup into the dolphin tube.

If the volume of your trimmed gel slice is too great to fit into one filter cup, then usetwo or more and pool the recovered samples ant the end of the protocol.

4. Place the Quantum Prep Freeze 'N Squeeze DNA Gel Extraction Spin Column (filter cup nested with dolphin tube) in a -20C freezer for five minutes.

5. Spin the sample at 13,000 x g for 3 minutes at room tempature.

6. Collect the purified DNA from the collection tube; the agarose debris will be retained within the filter cup of the Quantum Prep Freeze 'N Squeeze DNA Gel Extraction Spin Column. The DNA is read to use for PCR, ligations, labeling or other enzymatic reactions. Ethanol precipitation is recommeded for applications requiring a more concentrated sample and will also have the effect of further purifying the sample.

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