Detailed chromosomal and molecular genetic analysis of single cells by whole genome amplification and comparative genomic hybridisation

被引:208
作者
Wells, D
Sherlock, JK
Handyside, AH
Delhanty, JDA
机构
[1] Univ London Univ Coll, Galton Lab, Human Genet Unit, London NW1 2HE, England
[2] St Thomas Hosp, Dept Obstet & Gynaecol, London SE1 7EH, England
基金
英国惠康基金;
关键词
D O I
10.1093/nar/27.4.1214
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Molecular genetic analysis of isolated single cells and other minute DNA samples is limited because there is insufficient DNA to perform more than one independent PCR amplification. One solution to this problem is to first amplify the entire genome, thus providing enough DNA for numerous subsequent PCRs. In this study we have investigated four different methods of whole genome amplification performed on single cells, and have identified a protocol that generates sufficient quantities of DNA for comparative genomic hybridisation (CGH) as well as more than 90 independent amplification reactions. Thus, numerous specific loci and the copy number of every chromosome can be assessed in a single cell. We report here the first reliable application of CGH to single cells from human preimplantation embryos (blastomeres) and to single fibroblasts, buccal cells and amniocytes.
引用
收藏
页码:1214 / 1218
页数:5
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