Two efficient polymeric chemical platforms for oligonucleotide microarray preparation.
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.
Consolandi
Clarissa
C
Dipartimento di Scienze e Tecnologie Biomediche, Università degli Studi di Milano, LITA, Via Fratelli Cervi 93, 20090 Segrate Mi, Italy.
Castiglioni
Bianca
B
Bordoni
Roberta
R
Busti
Elena
E
Battaglia
Cristina
C
Bernardi
Luigi Rossi
LR
De Bellis
Gianluca
G
D000178:Acrylamides / Q000737:chemistry; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D004252:DNA Mutational Analysis / Q000379:methods; D019398:Genes, BRCA1; D005898:Glass / Q000737:chemistry*; D006801:Humans; D015394:Molecular Structure; D009154:Mutation / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes / Q000737:chemistry* / Q000235:genetics; D011108:Polymers / Q000737:chemistry*; D011437:Propylamines; D012821:Silanes / Q000737:chemistry*
D000178:Acrylamides; D015345:Oligonucleotide Probes; D011108:Polymers; D011437:Propylamines; D012821:Silanes; C431184:poly(acrylamide-co-acrylic acid); C477625:amino-propyl-triethoxysilane