Jacob Anette, Brandt Ole, Stephan Achim, Hoheisel Jörg D
Functional Genome Analysis, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Methods Mol Biol. 2004;283:283-93. doi: 10.1385/1-59259-813-7:283.
A fast and economical procedure for the production of peptide nucleic acid (PNA) microarrays is presented. PNA oligomers are synthesized in a fully automatic manner in 96-well plates using standard Fmoc chemistry. Subsequently, the oligomers are released from the support and spotted onto glass or silicone slides, which were activated by succinimidyl ester. This process allows for a concomitant purification of the oligomers directly on the chip surface. Although the terminal primary amino groups of the full-length products bind selectively to this surface, none of the byproducts of synthesis, such as truncated sequences or cleaved side chain protection groups, will bind and are therefore washed away. In this chapter, protocols are presented for the whole production process as well as sample hybridization.
介绍了一种快速且经济的肽核酸（PNA）微阵列生产方法。使用标准的芴甲氧羰基（Fmoc）化学方法在96孔板中以全自动方式合成PNA寡聚物。随后，将寡聚物从载体上释放并点样到经琥珀酰亚胺酯活化的玻璃或硅片上。该过程允许在芯片表面直接对寡聚物进行同步纯化。尽管全长产物的末端伯氨基会选择性地结合到该表面，但合成的副产物，如截短的序列或裂解的侧链保护基团，均不会结合，因此会被洗去。在本章中，给出了整个生产过程以及样品杂交的方案。
