How to keep microarray hybridization chambers from drying? - (Aug/21/2006 )

First, let me provide some background. I am using the Corning hybridization chambers for hybridizing microarrays. In each well of the chamber I load 15 ul water to provide humidity. I incubate my first hybridization (which is in an SDS-based buffer) overnight submerged in a 55 C waterbath. I can't use a formamide-based buffer because this causes a high background with the slides I am using (aldehyde-coated).

My problem is after an overnight incubation my hybridization chambers are drying out and my slides are drying. Does anyone have ideas or know of ways to maintain humidity with the procedure I have explained above? Is there a way I can modify my chambers?

-Zach

Put the whole thing in a box, add some wet paper towel around insdie the box, seal it.

Hi Zaprat,
in our case we are using manually hybridization in corning chamber , using formamide containing buffure. In our case we using torento spoted slide, we incubating hybridization slide at 42 for 18-20h not more than 20h. after three washing and drying slides by spining at 2000g for 5 min in 50 ml centrifuge tube we not geting any background i think its also work in your case you can try for this ....................for protocole i can provide for this experiment.
all the best and good luck.
awadh

hi zpratt,

that's weird, we also use the Corning hybridization chambers, and a urea based hybmix,
sometimes O/N at 65 degrees, but no drying,
we use 2 times 15µl hyb buffer to provide humidity and 80µl under the coverslip without probems,

- but what genehunter-1 is saying works also offcourse,
- you can also just seal your coverslip with glue (resistant against 55degrees)

good luck
A