Peter Hauser (last update 12.3.2002)

according to http://cmgm.stanford.edu/pbrown/protocols/index.html

Procedures

• Mark boundaries of array on the back of the slide using the diamond scriber
• Caveat: Array blocks become invisible after post-processing!
• Fill bottom of humid chamber with 1X SSC
• Place the slides upside-down on the humid chamber and close the lid
• Let the spots re-hydrate until they glisten (5 to 15 min, you may incubate shorter using higher temperature)
• Snap-dry the arrays (1-3 sec) on an inverted heat-block at 75°
• Place the arrays into slide rack
• Pipette 15 ml of the sodium borate solution into a 50 ml Falcon tube

Blocking solution:

• Dissolve 6 g succinic anhydride in approximately 325-350 ml 1-Methyl-2-pyrrolidinone
• Immediately after the succinic anhydride has dissolved, add 15 ml sodium borate solution and mix thoroughly
• Immediately after both solutions are mixed, pour solution into an empty slide chamber
• Plunge slide rack rapidly and evenly in solution and shake vigorously up and down for few seconds
• Mix on an orbital shaker for 15-20 min
• Meanwhile heat water in another slide chamber to boiling
• Gently plunge the slide rack in 95°C water (just stopped boiling) for 2 min
• Plunge slide rack 4-5 times into 95% ethanol
• Centrifuge slides (e.g. in 50 ml Falcon tubes, leave the tubes open) at 600 rpm for 3 min
• Use arrays or store them in a slide box

Materials: