Robust and efficient synthetic method for forming DNA microarrays
- 1 November 2001
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 29 (21) , 107e-107
- https://doi.org/10.1093/nar/29.21.e107
Abstract
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.Keywords
This publication has 31 references indexed in Scilit:
- DNA sequence analysis by hybridization with oligonucleotide microchips: MALDI mass spectrometry identification of 5mers contiguously stacked to microchip oligonucleotidesNucleic Acids Research, 2000
- Versatile derivatisation of solid support media for covalent bonding on DNA-microchipsNucleic Acids Research, 1999
- Light-Directed, Spatially Addressable Parallel Chemical SynthesisScience, 1991
- The structural gene for a phosphorus-repressible phosphate permease in Neurospora crassa can complement a mutation in positive regulatory gene nuc-1.Molecular and Cellular Biology, 1988
- Cloning of the am (glutamate dehydrogenase) gene of Neurospora crassa through the use of a synthetic DNA probeGene, 1982
- Electrophoretic transfer of DNA, RNA and protein onto diazobenzyloxymethyl (DBM) - paperNucleic Acids Research, 1980
- Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate.Proceedings of the National Academy of Sciences, 1979
- Quantitative analysis of specific labelled RNA's using DNA covalently linked to diazobenzyloxy-methyl-paperNucleic Acids Research, 1979
- Transfer of small DNA fragments from polyacrylamide gels to diazobenzyloxymethyl-paper and detection by hybridization with DNA probesBiochemical and Biophysical Research Communications, 1978
- Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes.Proceedings of the National Academy of Sciences, 1977