Detailed chromosomal and molecular genetic analysis of single cells by whole genome amplification and comparative genomic hybridisation
D Wells, JK Sherlock, JDA Delhanty… - Nucleic acids …, 1999 - academic.oup.com
D Wells, JK Sherlock, JDA Delhanty, AH Handyside
Nucleic acids research, 1999•academic.oup.comMolecular 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 …
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 …
Abstract
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.
Oxford University Press