Washing of microarrays?

[Peter Frank]

Hi,

I am preparing an oligonucleotide microarray, which is pretty similar
to doing a Northern blot. The microarray membrane is contained in a
small tube. Hybridization is done in a hybridizaton oven under
rotation at 7 rpm, which is within the recommended speed range.
However, the washing steps are to be performed at 20-30 rpm for around
15 min. Unfortunately, our hybridization oven only runs at 7 rpm,
other settings are not possible.
Based on your experience with Northern blots or microarrays, how would
you modify the washing steps if you could only do the washing steps at
7 rpm instead of 20-30 rpm? (After all, there are principles of
kinetics and equilibria behind all this.)

Peter

[Jose de las Heras]

Post by Peter Frank
Hi,
I am preparing an oligonucleotide microarray, which is pretty similar
to doing a Northern blot. The microarray membrane is contained in a
small tube. Hybridization is done in a hybridizaton oven under
rotation at 7 rpm, which is within the recommended speed range.
However, the washing steps are to be performed at 20-30 rpm for around
15 min. Unfortunately, our hybridization oven only runs at 7 rpm,
other settings are not possible.
Based on your experience with Northern blots or microarrays, how would
you modify the washing steps if you could only do the washing steps at
7 rpm instead of 20-30 rpm? (After all, there are principles of
kinetics and equilibria behind all this.)
Peter

it may sound too simple, but changing washing solution often will probably
be enough to ensure a good wash.

Jose