Antson Dan-Oscar, Mendel-Hartvig Maritha, Landegren Ulf, Nilsson Mats
The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, SE-751 85 Uppsala, Sweden.
Eur J Hum Genet. 2003 May;11(5):357-63. doi: 10.1038/sj.ejhg.5200966.
Conventional cytogenetic techniques can distinguish homologous chromosomes in a qualitative manner based upon obvious morphological features or using in situ hybridization methods that yield qualitative data. We have developed a method for quantitative genotyping of single-nucleotide variants in situ using circularizable DNA probes, so-called padlock probes, targeting two different alpha satellite repeat variants present in human chromosome 7 centromeres, and a single-nucleotide variation in alpha satellite repeats on human chromosome 15 centromeres. By using these PCR-generated padlock probes, we could quantitatively distinguish homologous chromosomes and follow the transmission of the chromosomes by in situ analysis during three consecutive generations.
传统的细胞遗传学技术可以根据明显的形态特征以定性方式区分同源染色体，或者使用能产生定性数据的原位杂交方法。我们开发了一种原位定量基因分型单核苷酸变异的方法，该方法使用可环化的DNA探针（即所谓的锁式探针），靶向人类7号染色体着丝粒中存在的两种不同的α卫星重复变异体，以及人类15号染色体着丝粒上α卫星重复序列中的一个单核苷酸变异。通过使用这些PCR生成的锁式探针，我们可以定量区分同源染色体，并通过连续三代的原位分析追踪染色体的传递情况。
