DNA Purification - GeneClean

Jeff Lawrence

This protocol employs the GeneClean Kit.

1. Electrophorese DNA fragment in an agarose gel using a TAE buffer system. Excise DNA slice in a minimal volume of agarose.

2. Weight the agarose slice. Aliquot 400 mg pieces into 1.5 mL microcentrifuge tubes.

3. Add 2.5 volumes of the NaI Solution. Incubate at 55° for 5 min to dissolve agarose.

4. Vortex the GlassMilk Solution to homogeneity.

5. Add GlassMilk Solution as follows:

a) Add 5 µL for 5 mg DNA or less.
b) Add 1 µL/mg DNA for DNA amounts > 5mg

6. Mix by inversion. Incubate on ice for 5 min.

7. Centrifuge for 5 sec at maximal speed at 4°. Discard supernatant.

8. Add 500 µL NEW Solution. Resuspend the pellet. Centrifuge for 5 sec at maximal speed. Discard the supernatant.

9. Repeat step (8) an additional 2 times.

10. Carefully remove the last of the NEW Solution with a pipet tip.

11. Resuspend the pellet in 10 µL TE. Incubate at 55° for 3 min.

12. Centrifuge for 30 sec at 22°.

13. Carefully remove the DNA-bearing supernatant from the pellet. Repeat elution for an additional 15% recovery.

Last Update: Thursday, 12-Dec-2024 00:55:23 UTC

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Eric Kofoid eckofoid at ucdavis.edu