SCHOTT Nexterion Slide A Protocol

Protocol
Short Version
Technical Instructions for Spotting Microarrays

PRODUCT

Nexterion Slide A is an activated glass slide in the standard size 75.5 mm x 25 mm x 1.0
mm with a multi-amino-silane* surface coating for efficient binding of cDNA molecules, PCR
products and longer oligonucleotides (size ≥ 50 mers).
*patent pending

DURABILITY AND STORAGE

Use by shelf-life date if stored in sealed condition. Open and use the slides in a clean
environment to avoid particle build-up on the printing surface. Avoid direct contact with the
printing surface to minimize contamination and abrasion of the coated surface. After opening
the package, store the remaining slides in a dust-free, light protected, and dry area at room
temperature and use within two weeks.

ARRAY PRINTING

1. Mix equal amounts of oligonucleotide probe or PCR product and 50% DMSO to obtain
a minimum final probe concentration of 20 µM for oligos, or 0.3 mg/ml for PCR
products. For smaller spot sizes, 3X SSC can be used as a printing buffer.
2. Transfer an appropriate volume of probes to a microtiter plate.
3. Set up the arrayer according to the manufacturer's recommendation and print slides at
40-50% relative humidity.

DNA IMMOBILIZATION

1. Rehydrate slides over a boiling water bath for 2-3 seconds, and then snap-dry for 10
seconds at 85°C on a hot plate.
2. UV cross link at 600 mJ for PCR products or 800 mJ for oligonucleotides.
3. Proceed immediately to prehybridization (washing and blocking) or store the arrays in
a desiccator for no more than two weeks.
<OPTIONAL> ARRAY DENATURATION (only for PCR products)
1. 1 x 30 sec in 0.1% SDS at room temperature.
2. 1 x 3 min in deionised H
3. 1 x 2 min 70% ethanol.
4. Spin-dry slides immediately at 200 x g for 5 min.

WASHING AND BLOCKING

1. 1 x 60 min Prehybridization Buffer (5X SSC, 0.1% SDS, 1.0% BSA, 50% formamide,
0.01% salmon sperm DNA) at 42°C.
2. 5 x deionised H
3. Spin-dry slides immediately.
O at 95°C.
2
O at room temperature.
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Nexterion™ Slide A
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