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Slide PreparationThe slide surface, the spotting buffer, the print head, and the spotting pins are critical components for reproducible, high-quality microarray printing. The laboratory staff at the Bauer Core has tested different spotting pins, spotting buffers, and slide coatings and can advise users as to the best combination for their application. The type of slide to use depends on the chemistry of the products to be printed. When printing amine-modified oligonucleotides, one should use either aldehyde- or epoxy-coated slides, which will react with the amine group on the oligonucleotide. These slides are available from several vendors at costs ranging from $6 to $11 per slide. When printing non-modified oligonucleotides or PCR products one should use either poly-lysine or aminosilane-coated slides. The coating on these slides gives them a positive charge which enables them to bind negatively charged DNA. Poly-lysine slides are available commercially, but are easy to make oneself at a fraction of the cost. The protocol for coating slides with poly-L-lysine takes two days, with about four hours of hands-on time. The core staff can provide training on how to make these slides. The reagents to coat 180 slides cost $160.18. One can coat fewer slides at a time, resulting in a lower reagent cost (although it is useful to have extra slides on hand for test prints). These slides should be aged for two weeks before use and used within two months of coating. Please direct questions about slide coating to Christian Daly (cdaly@cgr.harvard.edu). Poly-Lysine coating protocol (MS Word) |
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