Epplen C, Epplen J T
Molekulare Humangenetik, MA, Ruhr-Universität, Bochum, Germany.
Chromosome Res. 1995 Mar;3(2):131-4. doi: 10.1007/BF00710675.
In order to exclude inadvertent mixing of slides carrying metaphase spreads, chromosomal DNA was re-extracted several months after routine slide preparation (hypotonic treatment, spreading, fixation, staining, embedding and microscopic inspection) in different laboratories. The DNA yield was largely dependent on the number of cells originally fixed and the batch of the embedding material. Average fragment sizes ranged below 200 nucleotides. The polymerase chain reaction systems for DNA amplification included several polymorphic microsatellite loci specifically selected for short amplification products. Reidentification with a high probability for identity was possible by comparison with microsatellite alleles obtained from the peripheral blood of the individuals investigated. Possible applications, the differentiation potential and the limitations of the methodology are discussed.
为了排除携带中期染色体铺展的玻片意外混淆，在不同实验室进行常规玻片制备（低渗处理、铺展、固定、染色、包埋和显微镜检查）数月后，重新提取了染色体DNA。DNA产量在很大程度上取决于最初固定的细胞数量和包埋材料的批次。平均片段大小在200个核苷酸以下。用于DNA扩增的聚合酶链反应系统包括几个专门为短扩增产物选择的多态性微卫星位点。通过与从被调查个体外周血获得的微卫星等位基因进行比较，有可能以高概率进行身份重新鉴定。讨论了该方法的可能应用、分化潜力和局限性。
