Investigation of the multiple anchors approach in oligonucleotide microarray preparation using linear and stem-loop structured probes. (Articolo in rivista)

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Label
  • Investigation of the multiple anchors approach in oligonucleotide microarray preparation using linear and stem-loop structured probes. (Articolo in rivista) (literal)
Anno
  • 2002-01-01T00:00:00+01:00 (literal)
Alternative label
  • Bordoni R., Consolandi C., Castiglioni B., Busti E., Bernardi L.R., Battaglia C., De Bellis G. (2002)
    Investigation of the multiple anchors approach in oligonucleotide microarray preparation using linear and stem-loop structured probes.
    in Nucleic acids research
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • Bordoni R., Consolandi C., Castiglioni B., Busti E., Bernardi L.R., Battaglia C., De Bellis G. (literal)
Pagina inizio
  • E34 (literal)
Pagina fine
  • E40 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume
  • 30 (literal)
Rivista
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#note
  • IMPACT FACTOR 7.051 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#descrizioneSinteticaDelProdotto
  • Si tratta della messa a punto e validazione di una piattaforma chimica originale per la preparazione di DNA chip da utilizzarsi per l'analisi di mutazioni geniche. (literal)
Note
  • ISI Web of Science (WOS) (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
  • 1 CNR (ITB); 2 CNR (IBBA); 3 UNIMI (literal)
Titolo
  • Investigation of the multiple anchors approach in oligonucleotide microarray preparation using linear and stem-loop structured probes. (literal)
Abstract
  • Enzyme-mediated reactions are a useful tool in mutation detection when using a microarray format. Discriminating probes attached to the surface of a DNA chip have to be accessible to target DNA and to the enzyme (ligase or polymerase) that catalyses the formation of a new phosphodiester bond. This requires an appropriate chemical platform. Recently, an oligonucleotide hairpin architecture incorporating multiple phosphorothioate moieties along the loop has been proposed as an effective approach to solid-phase minisequencing. We have explored in depth several variables (stem length, number of phosphorothioates, stem-loop architecture versus linear structure) involved in this strategy by using a solid-phase ligation reaction. Microarrays were fabricated either from aminosilyl-modified glass or from aminated polymeric surfaces made of poly-lysine. Both platforms were bromoacetylated and reacted with thiophosphorylated oligonucleotides. The resulting microarrays were tested using either a synthetic template or a PCR-amplified 16S rRNA genomic region as the target sequence. Our results confirm the robustness of the proposed chemistry. We extend its range of application to solid-phase ligation, demonstrating the effectiveness of multiple anchors and suggest that linear oligonucleotides incorporating multiple phosphorothioates are equivalent to their hairpin-structured counterparts. (literal)
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