Search

An engineered polymer support 5 has been prepared for the solid-phase assembly of 3'-carboxyalkyl-modified oligonucleotides using commonly available reagents. A two-step deprotection procedure resulted in the quantitative cleavage of oligonucleotides from the support and removal of the protecting groups from phosphodiesters and exocyclic amino groups of the nucleic bases. The fully deprotected oligomers, obtained in high yield, were desalted and analyzed on RP-HPLC. After characterization by MALDI-TOF, these carboxyalkylated oligonucleotides were immobilized onto the epoxy-functionalized glass microslides to prepare biochips. The performance of these biochips was evaluated under different sets of conditions and then successfully validated by the detection of base mismatches and human infectious disease, bacterial meningitis, caused by N. meningitidis.

A new universal reagent, 2-O-[2-(4,4'-dimethoxytrityloxyethyl)]-hydroxy acetaldehyde (DEA), has been synthesized and used for the estimation of surface-bound aminoalkyl, aminooxyalkyl, hydrazinyl, and semicarbazide functions. The reaction completes in just 10 min in the case of aminoalkylated supports and 30 min in hydrazinyl supports, whereas it takes approximately 60 min in both aminooxyalkylated and semicarbazide-modified polymer supports. DEA-treated supports, including glass slides and PP films on exposure to acid, liberates 4,4'-dimethoxytrityl cation, which was measured spectrophotometrically to estimate these functionalities. The method estimates accessible functional groups, useful for calculating the quantity of the ligands to be immobilized.