DNA Purification - Wizard Columns

Jeff Lawrence

1. Add 100 µL of Extraction Buffer to liquid sample; mix well.

2. Add 1 µL of Beads. Vortex well three times for 5 sec, waiting 15 sec between repetitions.

3. Transfer solution to 3 mL syringe and slowly (2 drops/sec) apply to column.

4. Add 2 mL 80% Isopropanol to syringe and slowly apply to column.

5. Place column on a 1.5 mL Microfuge tube. Spin for 1 min.

6. Transfer column to a fresh tube. Add 50 µL TE to column. Wait 1 min.

7. Spin column in tube for 1 min. Discard column.

8. For Low-Melting-Point agarose, heat in extraction buffer at 42° until melted. Proceed as above.

9. For large (> 3 kb) fragments, elute DNA with TE at 65°.

Last Update: Thursday, 12-Dec-2024 00:55:24 UTC

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Eric Kofoid eckofoid at ucdavis.edu