Single DNA marker generated by “YAC-Alu PCR” that is end-specific

Abstract

SummaryA simple strategy for the rapid preparation of an end-specific linking-DNA probe from the YAC-human chromosome 21 DNA recombinant clone and the characterization of this single DNA probe are described. Synthetic oligodeoxynucleotide primers, based on the consensus Alu sequence, and the Sup4 DNA fragment in the YAC arms were used to amplify end-specific DNA sequences by the polymerase chain reaction (PCR) for screening of the linking YAC recombinant clones ("YAC-Alu PCR"). Nucleotide sequencing of the product of PCR from human genomic DNA in a YAC insert confirmed the boundary between the vector and the insert and the presence of the 3'-end Alu-like structure. The probe R1, prepared by "YAC-Alu PCR" amplification, was assigned to chromosome 21 by Southern hybridization of somatic cell hybrid DNAs. In situ hybridization allowed localization of the R1 DNA probe to the human chromosome 21q21-q22.1 region. Thus, this approach has significant advantages not only for isolation of a single DNA probe specific for human chromosome 21 but also for the screening of YAC linking recombinant clones for mapping of the human genome.

Yeast artificil chromosome (YAC) clones and sequence tagged site (STS) markers anchored at human chromosome 21

“…The total length of human DNA inserts was calculated about 6.6 Mb, which is equivalent to 16.5% that of the human chromosome 21. With these YAC clones anchoring at chromosome 21, we are now trying to isolate YAC linking clones to make a contig map of human chromosome 21 by means of "inverse-PCR" (Riley et al, 1990) or "YAC-Alu PCR" (Breukel et al, 1990;Cole et al, 1991;Tashiro et al, 1991).…”