Document Detail

Single-cell isolation from cell suspensions and whole genome amplification from single cells to provide templates for CGH analysis.
MedLine Citation:
PMID:  18079717     Owner:  NLM     Status:  MEDLINE    
A comprehensive genomic analysis of single cells is instrumental for numerous applications in tumor genetics, clinical diagnostics and forensic analyses. Here, we provide a protocol for single-cell isolation and whole genome amplification, which includes the following stages: preparation of single-cell suspensions from blood or bone marrow samples and cancer cell lines; their characterization on the basis of morphology, interphase fluorescent in situ hybridization pattern and antibody staining; isolation of single cells by either laser microdissection or micromanipulation; and unbiased amplification of single-cell genomes by either linker-adaptor PCR or GenomePlex library technology. This protocol provides a suitable template to screen for chromosomal copy number changes by conventional comparative genomic hybridization (CGH) or array CGH. Expected results include the generation of several micrograms of DNA from single cells, which can be used for CGH or other analyses, such as sequencing. Using linker-adaptor PCR or GenomePlex library technology, the protocol takes 72 or 30 h, respectively.
Jochen B Geigl; Michael R Speicher
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Nature protocols     Volume:  2     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2007  
Date Detail:
Created Date:  2007-12-14     Completed Date:  2008-02-19     Revised Date:  2008-03-24    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  3173-84     Citation Subset:  IM    
Institute of Human Genetics, Center for Applied Biomedicine, Medical University of Graz, Harrachgasse 21/8, A-8010 Graz, Austria.
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MeSH Terms
Cell Line, Tumor
Cell Separation / methods
Gene Library
Genomics / methods*
In Situ Hybridization, Fluorescence
Nucleic Acid Amplification Techniques
Oligonucleotide Array Sequence Analysis / methods

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