Modified Gel Extraction Protocol
Steps 1-4 with QIAquick® Gel Extraction Kit, steps 5-8 with Zymoclean™ Gel DNA Recovery Kit
- Excise the DNA fragment from the agarose gel with a clean, sharp scalpel.
- Weigh the gel slice in a colorless tube. Add 3 volumes Buffer QG to 1 volume gel (100 mg gel ~ 300 µL QG). The maximum amount of gel per spin column is 400 mg. For >2% agarose gels, add 6 volumes Buffer QG.
- Incubate at 50 °C for 10 min (or until the gel slice has completely dissolved). Vortex the tube every 2-3 min to help dissolve gel. After the gel slice has dissolved completely, check that the color of the mixture is yellow (similar to Buffer QG without dissolved agarose). If the color of the mixture is orange or violet, add 10 µL 3 M sodium acetate, pH 5.0, and mix. The mixture turns yellow.
- Add 1 volume isopropanol to the sample and mix.
- Load the dissolved solution into a Zymo-Spin Column in a Collection Tube.
- Centrifuge at full speed (≥10,000 x g) for 30 seconds. Discard the flow through.
- Add 200 µL of DNA Wash Buffer to the column and centrifuge for 30 seconds. Repeat the wash step.
- Place the Zymo-Spin Column into a new 1.5 mL tube. Dry the column at 50 °C for 10 min. Add ≥ 6 µL of DNA Elution Buffer or warm water directly to the column matrix and spin to elute the DNA.