期刊
BIOCONJUGATE CHEMISTRY
卷 25, 期 3, 页码 618-627出版社
AMER CHEMICAL SOC
DOI: 10.1021/bc500033d
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资金
- Ministerio de Ciencia e Innovacion [CTQ2010-15943]
- Generalitat Valenciana [PROMETEO/2010/008, ACOMP/2012/158]
A rapid strategy for the covalent immobilization of DNA onto silicon-based materials using the UV-initiated radical thiol-ene reaction is presented in this study. Following this approach, thiol- and alkene-modified oligonucleotide probes were covalently attached in microarray format, reaching immobilization densities around 6 pmol-cm(-2). The developed methodology presents the advantages of spatially controlled probe anchoring (using a photomask), direct attachment without using cross-linkers (one-pot fashion), and short irradiation times (20 min). Using the described strategy, hybridization efficiencies up to 65% with full complementary strands were reached. The approach was evaluated by scoring single-base pair mismatches with discrimination ratios around 15. Moreover, the efficacy of the proposed DNA detection scheme is further demonstrated through the assay on a genomic target of bacterial Escherichia coli.
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