DNA Hybridization - Filter Rinsing

Jeff Lawrence

1. Remove filter from hybridization bag and place in 10 mL of Wash Solution:

	1 mM	Tris, pH 8.0	100 mL	1 M Tris, pH 8.0
	1 %	Sarkosyl	10 mL	10 % Sarkosyl
				90 mL	ddH₂O

2. Rinse filter for 10 sec to remove excess hybridization fluid. Remove solution with a pipet and discard.

3. Add 90 mL Wash Solution. Incubate at 22° for 15 min with agitation.

4. Decant wash solution. Add 250 mL Rinse Solution:

	1 mM	Tris, pH 8.0	1 mL	1 M Tris, pH 8.0
				999 mL	ddH₂O

5. Rinse at 22° for 5 min with agitation. Monitor radioactivity of filter.

6. Repeat rinses until background radioactivity is removed.

7. Wrap filter in plastic wrap and expose to X-ray film.

Last Update: Thursday, 12-Dec-2024 00:55:21 UTC

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Eric Kofoid eckofoid at ucdavis.edu