In situ synthesis of oligonucleotide arrays by using the molecular stamp method

This work developed a new technique for in situ synthesis of oligonucleotide arrays on glass surfaces. In this method, based on the standard phosphoramidites chemistry protocol, the coupling was conducted through contacting glass surfaces with a set of molecular stamps on which surfaces spread nucleoside monomer and tetrazole mixed acetonitrile solution. It was shown that 20-mer oligonucleotide on the glass slide was successfully synthesized using the modified polydimethylsiloxane (PDMS) stamps and showed the high synthetic efficiency. An effective method has been used to eliminate residual reactive nucleosides on chip with small molecules containing hydroxyl group. A specific oligonucleotide arrays of four probes including matched and mismatched with the target sequence was fabricated to identify the perfect match and mismatch sequences. It indicated that the perfect match and mismatch probes hybridization fluorescent signals had clearly difference, and may be used to identify rapidly screen single-nucleotide polymorphisms.