The "Clickable" Method for Oligonucleotide Immobilization Onto Azide-Functionalized Microarrays

Methods Mol Biol. 2016:1368:25-36. doi: 10.1007/978-1-4939-3136-1_3.

Abstract

The DNA microarray technique was supposed to help identifying and analyzing the expression level of tens of thousands of genes in the whole genome. But there is a serious problem concerning fabrication of the microarrays by chemical synthesis, such as specific and efficient linking of probes to a solid support. Therefore, we reckon that applying "click" chemistry to covalently anchor oligonucleotides on chemically modified supports may help construct microarrays in applications such as gene identification. Silanization of the glass support with organofunctional silane makes it possible to link azide groups on glass surface and the nucleic acid probe that is equipped with a pentynyl group. This is followed by direct spotting of the nucleic acid on the azide-modified glass support in the presence of copper ions, and this is a frequently applied method of "click" chemistry.

Keywords: Click chemistry; DNA; Fluorescence; Hybridization; Microarrays; Oligonucleotides; Phosphoramidite; Silanization.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ascorbic Acid / chemistry
  • Azides / chemistry*
  • Click Chemistry / methods*
  • Copper Sulfate / chemistry
  • Glass / chemistry
  • Glycerol / chemistry
  • Nucleic Acid Hybridization
  • Oligodeoxyribonucleotides / chemical synthesis
  • Oligodeoxyribonucleotides / chemistry
  • Oligonucleotide Array Sequence Analysis / methods*
  • Oligonucleotides / chemistry*
  • Silanes / chemistry

Substances

  • Azides
  • Oligodeoxyribonucleotides
  • Oligonucleotides
  • Silanes
  • Copper Sulfate
  • Glycerol
  • Ascorbic Acid