In situ oligonucleotide synthesis on carbon materials: stable substrates for microarray fabrication.
Glass has become the standard substrate for the preparation of DNA arrays. Typically, glass is modified using silane chemistries to provide an appropriate functional group for nucleic acid synthesis or oligonucleotide immobilization. We have found substantial issues with the stability of these surfaces as manifested in the unwanted release of oligomers from the surface when incubated in aqueous buffers at moderate temperatures. To address this issue, we have explored the use of carbon-based substrates. Here, we demonstrate in situ synthesis of oligonucleotide probes on carbon-based substrates using light-directed photolithographic phosphoramidite chemistry and evaluate the stabilities of the resultant DNA arrays compared to those fabricated on silanized glass slides. DNA arrays on carbon-based substrates are substantially more stable than arrays prepared on glass. This superior stability enables the use of high-density DNA arrays for applications involving high temperatures, basic conditions, or where serial hybridization and dehybridization is desired.
Phillips
Margaret F
MF
Department of Chemistry, University of Wisconsin - Madison, 1101 University Ave. Madison, Madison, WI 53706, USA.
Lockett
Matthew R
MR
Rodesch
Matthew J
MJ
Shortreed
Michael R
MR
Cerrina
Franco
F
Smith
Lloyd M
LM
D002244:Carbon / Q000737:chemistry*; D018130:Diamond / Q000737:chemistry; D005456:Fluorescent Dyes / Q000032:analysis; D005898:Glass / Q000737:chemistry; D008856:Microscopy, Fluorescence; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes / Q000138:chemical synthesis* / Q000737:chemistry; D016133:Polymerase Chain Reaction; D013696:Temperature
D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D002244:Carbon; D018130:Diamond