Two efficient polymeric chemical platforms for oligonucleotide microarray preparation

Nucleosides Nucleotides Nucleic Acids. 2002;21(8-9):561-80. doi: 10.1081/NCN-120015069.

Abstract

In this report we describe two robust procedures for oligonucleotide microarray preparation based on polymeric coatings. The proposed chemical approaches include: 1) a glass functionalisation step with appropriate silanes (gamma-aminopropyltriethoxysilane-APTES or 3-glycidoxypropyltrimethoxysilane-GOPS), 2) a coating step using polymers (poly-L-Lysine or poly(acrylic acid-co-acrylamide) copolymer) covalently bound to the modified glass and 3) a surface activation step to allow for the attachment of amino-modified oligonucleotides. Results obtained using these chemistries in oligo microarray preparation show: 1) an overall high loading capacity and availability to hybridisation against targets, 2) a good uniformity, 3) resistance to consecutive probing/ stripping cycles, 4) stability to thermal cycles, 5) effectiveness in hybridisation-mediated mutation detection procedures and 6) the possibility to perform enzymatic reactions, such as ligation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylamides / chemistry
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • DNA Mutational Analysis / methods
  • Genes, BRCA1
  • Glass / chemistry*
  • Humans
  • Molecular Structure
  • Mutation / genetics
  • Oligonucleotide Array Sequence Analysis*
  • Oligonucleotide Probes / chemistry*
  • Oligonucleotide Probes / genetics
  • Polymers / chemistry*
  • Propylamines
  • Silanes / chemistry*

Substances

  • Acrylamides
  • Oligonucleotide Probes
  • Polymers
  • Propylamines
  • Silanes
  • poly(acrylamide-co-acrylic acid)
  • amino-propyl-triethoxysilane