Document Detail

Measurement of single-cell gene expression using capillary electrophoresis.
MedLine Citation:
PMID:  11774920     Owner:  NLM     Status:  MEDLINE    
Capillary electrophoresis with laser-induced fluorescence detection was used to monitor gene expression in individual mammalian cells using the reverse transcriptasepolymerase chain reaction. Specifically, beta-actin expression in single LNCaP (prostate cancer) cells was measured. A sieving matrix containing hydroxypropyl methyl cellulose was used to effect size-based separation. Ethidium bromide fluorescence of the product DNA was used as the detection scheme and yielded excellent sensitivity. The beta-actin product, resulting from an individual cell lysed by a freeze-thaw method, gave an average signal-to-noise ratio (S/N) of 77+/-27 (n = 2). Chemical lysis of a single cell, using a dilute solution of SDS, gave a S/N of 26+/-2 (n = 2), roughly 3-fold lower than for freeze-thaw lysis. An initial detection limit (not considering fully optimized conditions) was calculated from an amplified cDNA standard to correspond to a concentration of approximately 133 starting molecules/nL (of beta-actin mRNA).
J L Zabzdyr; S J Lillard
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  73     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2001 Dec 
Date Detail:
Created Date:  2001-12-31     Completed Date:  2002-02-01     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5771-5     Citation Subset:  IM    
Department of Chemistry, University of California, Riverside 92521, USA.
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MeSH Terms
Electrophoresis, Capillary
Gene Expression*
Protein Biosynthesis
Proteins / analysis*,  genetics
Reverse Transcriptase Polymerase Chain Reaction
Spectrometry, Fluorescence
Tumor Cells, Cultured
Reg. No./Substance:

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