Array Stripping

9 October, 2000

1. In a Saran Wrap covered 11 x 7 in Pyrex dish bring approximately 200 ml stripping solution to a boil using microwave at power setting 10 [high] (2-3min).

2. Add the blot to the heated solution, re-cover with Saran Wrap, and microwave for 20 min at power level 3 (maintains stripping solution at a low boil, similar to defrost setting).

3. Drain the excess stripping solution and rinse array briefly in 2X SSPE.

4. Re-wrap the array (in Saran Wrap or by the alternative sheet protector method) and expose array to phosphor imaging screen for a length of time equal to the original experiment.

5. Greater than 95% of the signal should have been stripped. If significant signals persist, then repeat the stripping procedure once more. Always use fresh stripping solution.

6. If the array is stripped, proceed to a new hybridization experiment. Store unused arrays in Saran Wrap at 4oC.

Stripping Solution:

10mM Tris, pH 7.5/8.0 5mls of a 1M stock
1mM EDTA 1 ml of a 0.5 M stock
1% SDS 25mls of a 20% stock soln

Add 469 ml of H2O to make a total volume of 500 mls.
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OU Bioinformatics Core Facility @ Advanced Center for Genome Technology | Credits | updated:19 Oct 2005