DNA Purification - GeneClean
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, 19-Jun-2014 11:47:12 PDT
This page has been viewed 383 times.
eckofoid at ucdavis.edu