A structured chitosan-based platform for biomolecule attachment to solid surfaces: application to DNA microarray preparation.
A structured chemical platform based on chitosan, an amine-rich polysaccharide, is presented as an alternative chemistry to functionalize solid support (in this case, glass slides) for grafting biomolecules. This approach has been adopted for generating arrays using amino-modified oligonucleotides with two different lengths (25-mer and 70-mer) for different purposes. Results using these chitosan-activated surfaces indicate high oligonucleotide loading capacity, good availability to hybridization against targets, and effectiveness in enzyme-mediated single nucleotide polymorphism (SNP) detection procedures by DNA polymerase and DNA ligase enzymes with low background. Universal arrays have been prepared and extensively used with excellent results in different applications. The chitosan-treated surfaces were also evaluated for their performance in a gene expression experiment.
Istituto di Tecnologie Biomediche, Consiglio Nazionale delle Ricerche, LITA, via F.lli Cervi, 93 20090 Segrate (MI) Italy. clarissa.consolandi@itb.cnr.it
Clarissa
C
Consolandi
Marco
M
Severgnini
Bianca
B
Castiglioni
Roberta
R
Bordoni
Andrea
A
Frosini
Cristina
C
Battaglia
Luigi Rossi
LR
Bernardi
Gianluca De
GD
Bellis
D048271:Chitosan / Q000737:chemistry*; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D009841:Oligonucleotides / Q000737:chemistry*; D012313:RNA / Q000378:metabolism; D013499:Surface Properties
D009841:Oligonucleotides; D012313:RNA; D048271:Chitosan