Robust and efficient synthetic method for forming DNA microarrays.
The field of DNA microarray technology has necessitated the cooperative efforts of interdisciplinary scientific teams to achieve its primary goal of rapidly measuring global gene expression patterns. A collaborative effort was established to produce a chemically reactive surface on glass slide substrates to which unmodified DNA will covalently bind for improvement of cDNA microarray technology. Using the p-aminophenyl trimethoxysilane (ATMS)/diazotization chemistry that was developed, microarrays were fabricated and analyzed. This immobilization method produced uniform spots containing equivalent or greater amounts of DNA than commercially available immobilization techniques. In addition, hybridization analyses of microarrays made with ATMS/diazotization chemistry showed very sensitive detection of the target sequence, two to three orders of magnitude more sensitive than the commercial chemistries. Repeated stripping and re-hybridization of these slides showed that DNA loss was minimal, allowing multiple rounds of hybridization. Thus, the ATMS/diazotization chemistry facilitated covalent binding of unmodified DNA, and the reusable microarrays that were produced showed enhanced levels of hybridization and very low background fluorescence.
Dolan
P L
PL
Department of Biology, University of New Mexico, Albuquerque, NM 87131, USA.
Wu
Y
Y
Ista
L K
LK
Metzenberg
R L
RL
Nelson
M A
MA
Lopez
G P
GP
D000327:Adsorption; D002232:Carbocyanines / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000378:metabolism; D020869:Gene Expression Profiling / Q000379:methods; D005898:Glass; D009492:Neurospora crassa / Q000235:genetics; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011107:Polylysine / Q000378:metabolism; D015203:Reproducibility of Results; D012821:Silanes / Q000737:chemistry / Q000378:metabolism
D002232:Carbocyanines; D015342:DNA Probes; D004271:DNA, Fungal; D005456:Fluorescent Dyes; D012821:Silanes; C089187:cyanine dye 3; C085321:cyanine dye 5; D011107:Polylysine; D004247:DNA; C071717:trimethoxysilane