DNA Transfer - Electroblot


Jeff Lawrence

1. Prechill 2.5 L 0.5X TBE at 4° in the electroblot chamber.

2. Soak 2 sponges in 0.5X TBE. Prepare 3 Whatman 3MM sheet and 1 Nylon filter.

3. Run acrylamide gel until Bromphenol blue dye is running off. Remove 1 plate and trim excess acrylamide.

4. Wet the gel with water to pre-contract. Pick up gel on Whatman 3MM paper. Slice gel in half.

5. Place 1 piece of prewet Whatman 3MM on wet sponge. Roll out air bubbles.

6. Place the top half of the gel/Whatman on top and cover with Nylon.

7. Place the second piece of Whatman paper on top.

8. Place the bottom half of the gel/Whatman on top and cover with Nylon.

9. Place the third piece of Whatman paper on top. Cover all with second sponge.

10. Place all in clamp holder and immerse in prechilled buffer chamber.

11. Electroblot for 60 min at 50 V (500 mA).

12. Remove filter and airdry. Crosslink DNA by exposure to germicidal strength UV light for 5 min.

Last Update: Thursday, 19-Jun-2014 11:50:31 PDT
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Eric Kofoid eckofoid at ucdavis.edu