Lucito R, Nakimura M, West J A, Han Y, Chin K, Jensen K, McCombie R, Gray J W, Wigler M
Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA.
Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4487-92. doi: 10.1073/pnas.95.8.4487.
Analysis of the genetic changes in human tumors is often problematical because of the presence of normal stroma and the limited availability of pure tumor DNA. However, large amounts of highly reproducible "representations" of tumor and normal genomes can be made by PCR from nanogram amounts of restriction endonuclease cleaved DNA that has been ligated to oligonucleotide adaptors. We show here that representations are useful for many types of genetic analyses, including measuring relative gene copy number, loss of heterozygosity, and comparative genomic hybridization. Representations may be prepared even from sorted nuclei from fixed and archived tumor biopsies.
由于正常基质的存在以及纯肿瘤DNA的可得性有限，对人类肿瘤中的基因变化进行分析往往存在问题。然而，通过聚合酶链反应（PCR），可以从纳克量的经限制性内切酶切割并连接到寡核苷酸接头的DNA中，制备出大量高度可重复的肿瘤和正常基因组“代表性片段”。我们在此表明，这些代表性片段可用于多种类型的基因分析，包括测量相对基因拷贝数、杂合性缺失以及比较基因组杂交。甚至可以从固定存档的肿瘤活检组织中分拣出的细胞核制备代表性片段。
