The development of sequence-tagged sites for human chromosome 4

Abstract

As part of our efforts to construct a high-resolution physical map of human chromosome 4, we developed a systematic approach for efficiently generating large numbers of chromosome-specific sequence-tagged sites (STSs). In this paper, we describe how rate-limiting steps in our STS development were identified and overcome, and detail our current development strategy. We present information for 822 new human chromosome 4-specific STSs, including PCR amplification conditions and subchromosomal localization data, obtained by analysis of the STS with somatic cell hybrids containing different portions of human chromosome 4. Although most STSs presented here were developed from anonymous clones whose sequences were determined in this laboratory, several STSs were developed for genes and other DNA sequences that were previously mapped to chromosome 4. Our data indicate that the availability of DNA sequence for an STS locus, in addition to the sequences of the two PCR oligonucleotides, significantly increases the transfer of that STS by allowing investigators to select new oligonucleotides best suited to the standard conditions used in their laboratories.