@misc{oai:repo.qst.go.jp:00069676,
 author = {Michikawa, Yuichi and Noshiro, Katsuko and Suga, Tomo and Ishikawa, Atsuko and Shoji, Yoshimi and Iwakawa, Mayumi and Imai, Takashi and 道川 祐市 and 野代 勝子 and 菅 智 and 石川 敦子 and 荘司 好美 and 岩川 眞由美 and 今井 高志},
 month = {Dec},
 note = {OBJECTIVES: Haploid chromosomes distinguished by multiple SNP markers in human PTTG1 (Chromosome 5) and CD44 (Chromosome 11) genes have been reported to associate with the severity of adverse effects induced by radiation therapy of breast cancer patients. It is of interest to understand biological consequences of the heterozygotic state by functionally distinct, homologous haploid chromosomes within a nucleus of cells in the patients. To this end, diplotype of these chromosomes must be precisely determined to evaluate their clinical outcomes. 
\nMETHODS: In this study, a novel methodology of long-range (over 20 kb) haplotype determination involving sensitive amplification of single DNA molecules within agarose gels has been developed. Limited dilution of DNA molecules was performed in heated alkaline agarose solution, avoiding extensive shearing and aggregation during dilution process. Aliquoting the gel solution provided physical separation of DNA molecules derived from homologous chromosomes. The solidified agarose gel pieces were then treated by exogenously supplied Phi29 DNA polymerase and random hexamer oligonucleotides, yielding up to 120,000-fold multiple-displacement amplification of the gel-embedded DNA molecules. The amplified materials were recovered in solution as PCR-ready form by simple heating, making them conveniently available for further multiple locus genotyping. 
\nRESULTS: and CONCLUSIONS Feasibility of this methodology for isolating long-range single DNA molecules spanning the selected SNP markers (PTTG1: 20 kb, CD44: 80 kb) was further confirmed using DNAs derived from patients with heterozygotic state. Thus, the haploid chromosomes distinguished by the SNP markers of the above clinical radiation sensitivity-associated genes can now be successfully determined in the individual breast cancer patients., 48ｔｈ the American Society for Cell Biology Annual Meeting},
 title = {Long-range single DNA molecule haplotyping of human clinical radiation sensitivity-associated genes (PTTG1 and CD44)},
 year = {2008}
}