Remove chamber cover from probed array over garbage can as hybridization solution will drip.

Carefully remove any remaining adhesive material from the array with a razor blade.

Places arrays in Wheaton glass slide holder.

Wash in pre-warmed 2X SSC, 0.2% SDS washing solution at 65oC for 5 minutes.

(Every minute lift the tray up and down 2-3x to aid in washing)

Wash the array in 2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Wash the array in 0.2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Wash the array in 0.2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Air Dry the array.

11. Hybridization of Dendrimers (Cy3 and Cy5) to Array (NON-Automated version).

Place circular filter papers (about 3-4) into both reservoirs of pre-warmed incubation chamber.

Add 40-50 ul of 2X SSC onto each filter paper stack.

Place the air-dried array into the chamber. (DNA side up)

Add 2 ul of 65oC dendrimer mixture into 13 ul of 65oC 3DNA hybridization buffer and mix well.

(These values are for a 25 mm x 25 mm cover slip)

(Double the volumes for a 40 mm x 25 mm cover)

Place the 15 ul hybridization solution and dendrimers on the center of the array.

(Between the scratches made with the diamond pencil)

Carefully place the ethanol wiped and dried glass cover slip (Corning) on the drop of solution.

If necessary, remove bubbles by gentle tapping on glass with a pipette tip.

Assemble the incubation chamber cover on the base. (MAKE SURE GASKET IS SEATED)

Apply a binder clip at each end, and fold the metal hinges toward the middle.

Place the chamber and array at 65oC for 6 hours.

12. Washing unbound dendrimer from glass array(NON-Automated version).

Remove incubation chamber from 65oC incubator.

Carefully remove binder clips and cover from incubation chamber.

Places arrays in Wheaton glass slide holder.

Wash in pre-warmed 2X SSC, 0.2% SDS washing solution at 65oC for 10 minutes AFTER COVER

SLIP FALLS OFF!

(Every minute lift the tray up and down 2-3x to aid in washing)

(Make sure that the cover slip falls off, do not help in any other way)

Wash the array in 2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Wash the array in 0.2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Wash the array in 0.2X SSC, 0.2% SDS washing solution at RT for 2 minutes.

Air Dry the array.

Wipe the back of the array with a Kimwipe (70 % ETOH treated).

Scan the array.