High-fidelity digital hybridization screening.
We developed a highly efficient screening method for minimizing the hybridization of high-density replica (HDR) filters and for allowing simultaneous use of numerous oligonucleotide probes for STS markers. We designated this method "digital hybridization (DH) screening," in which a binary n-bit ID number is given to each probe, and a series of probe mixtures is prepared in an arranged combination. The matrix pattern between probe mixtures and hybridization signals determines the relation between a particular STS marker(s) and the corresponding DNA clone(s). Here, we describe a successful DH screening of over 15,000 human BAC clones with 126 STS marker probes with 7-bit ID numbers, which required only 8 sets of HDR filter hybridizations. Our results indicate that DH screening can be performed with more than 1000 STS marker probes with only 10-bit ID numbers. The DH screening method is convenient, economical, and of high fidelity, and thus it should facilitate the construction of sequence-ready DNA contigs for the human genome as well as for genomes of various species. The principle of DH screening has various applications to the biological sciences.
Asakawa
S
S
Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.
Shimizu
N
N
D000818:Animals; D002876:Chromosomes, Bacterial / Q000235:genetics; D003001:Cloning, Molecular; D015698:Genomic Library; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes; D016324:Sequence Tagged Sites
D015345:Oligonucleotide Probes