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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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