COMBINING THE PREPARATION OF OLIGONUCLEOTIDE ARRAYS AND SYNTHESIS OF HIGH-QUALITY PRIMERS

Based on the oligomer-chip technology, oligonucleotide arrays mere syn thesized directly on polypropylene sheets by a modified phosphoramidit e chemistry using beta-eliminating nucleobase-protecting groups in com bination with a succinate solid-phase linker. This method decouples th e oligonucleotide deprotection from the support cleavage procedure, in contrast to standard phosphoramidite chemistry. In addition to being reliable substrates for hybridization experiments, the arrays also ser ve as source for the isolation of individual oligonucleotides. Technic ally, this allowed for a direct control of the quality of the arrayed oligomers. The released compounds were sufficient in amount and purity to work without further purification in PCR and DNA-sequencing reacti ons, with the results being identical to controls with commercially ob tained primer molecules. Consequences for oligomer-chip hybridization procedures, the applicability of such hybrid-function arrays in, for e xample, diagnostics or comparative biology, and developments toward pa rallel primer synthesis are discussed. (C) 1996 Academic Press, Inc.