Issue August 2000

category image Volume 18
No. 1 (p 1-168)
August 2000
ISSN 0739-1102

Sequencing by Hybridization with the Generic 6-mer Oligonucleotide Microarray: An Advanced Scheme for Data Processing (p. 83-102)

DNA sequencing by hybridization was carried out with a microarray of all 46 = 4,096 hexadeoxyribonucleotides (the generic microchip). The oligonucleotides immobilized in 100 × 100 × 20-µm 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.

V. R. Chechetkin1,2
A. Y. Turygin1,2
D. Y. Proudnikov1,2
D. V. Prokopenko1
E. V. Kirillov1
A. D. Mirzabekov1,2*

Joint Human Genome Program:
1Engelhardt Institute of Molecular Biology
Russian Academy of Sciences
32 Vavilov Str.
117984 Moscow, Russia
2Biochip Technology Center
Argonne Nat. Lab.
9700 S. Cass Ave.
Argonne, IL 60439
*amir@everest.bim.anl.gov

Purchase Downloadable Full Text PDF of Articles

Corporate User

$100.00

University/Academic User

$50.00

Subscription is more cost effective than purchasing PDFs on-the-fly.  Click here for details.