Sequencing by hybridization with the generic 6-mer oligonucleotide microarray : an advanced scheme for data processing.
DNA sequencing by hybridization was carried out with a microarray of all 4{sup 6} = 4,096 hexadeoxyribonucleotides (the generic microchip). The oligonucleotides immobilized in 100 x 100 x 20-{mu}m polyacrylamide gel pads of the generic microchip were hybridized with fluorescently labeled ssDNA, providing perfect and mismatched duplexes. Melting curves were measured in parallel for all microchip duplexes with a fluorescence microscope equipped with CCD camera. This allowed us to discriminate the perfect duplexes formed by the oligonucleotides, which are complementary to the target DNA. The DNA sequence was reconstructed by overlapping the complementary oligonucleotide probes. We developed a data processing scheme to heighten the discrimination of perfect duplexes from mismatched ones. The procedure was united with a reconstruction of the DNA sequence. The scheme includes the proper definition of a discriminant signal, preprocessing, and the variational principle for the sequence indicator function. The effectiveness of the procedure was confirmed by sequencing, proofreading, and nucleotide polymorphism (mutation) analysis of 13 DNA fragments from 31 to 70 nucleotides long.
- Research Organization:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Sponsoring Organization:
- USDOE; FOR
- DOE Contract Number:
- DE-AC02-06CH11357
- OSTI ID:
- 942879
- Report Number(s):
- ANL/BTC/JA-35547; JBSDD6; TRN: US200923%%667
- Journal Information:
- J. Biomol. Struct. Dyn., Vol. 18, Issue 1 ; Aug. 2000; ISSN 0739-1102
- Country of Publication:
- United States
- Language:
- ENGLISH
Similar Records
PCR amplfication on a microarray of gel-immobilized oligonucleotides : detection of bacterial toxin- and drug-resistent genes and their mutations.
Parallel thermodynamic analysis of duplexes on oligodeoxyribonucleotide microchips.