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

This work developed a 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 the surfaces of which nucleoside monomer and tetrazole mixed acetonitrile solution were spread. It was shown that 20-mer oligonucleotide on the glass slide was successfully synthesized using the modified polydimethylsiloxane stamps and showed high synthetic efficiency. An effective method has been used to eliminate residual reactive nucleosides on chip with small molecules containing a hydroxyl group. Specific oligonucleotide arrays of four probes including matches and mismatches with the target sequence were fabricated to identify the perfect match and mismatch sequences. Results indicated that the perfect match and mismatch probes hybridized fluorescent signals had clear differences, and may be used to identify and rapidly screen single-nucleotide polymorphisms.