PMID: 11334978May 4, 2001Paper

Profiling of gene expression in individual hematopoietic cells by global mRNA amplification and slot blot analysis

Journal of Immunological Methods
K Theilgaard-MönchN Borregaard

Abstract

The pattern of expressed genes defines the structure and functional status of cells. Currently, most methods used in gene expression studies depend on large numbers of cells. Thus, their application may be hampered by the heterogeneity of cell populations, and by the low numbers of cells obtainable from in vivo sources. Such drawbacks may be overcome by methods suitable for the profiling of gene expression at the single cell level. We studied whether polymerase chain reaction (PCR) products synthesized from individual cells by global amplification of messenger RNA (mRNA) were suitable as probes for gene expression analysis. For this purpose, cells were subjected to reverse transcription and PCR using sequence independent primers (SIP RT-PCR). The resultant cDNA products were radiolabeled and hybridized to cDNA clones arrayed on a nylon membrane by vacuum slot blotting (a method referred to as slot blot analysis). The SIP RT-PCR procedure was reproducible and allowed the detection of twofold changes in input RNA copies per cell (range: 80-10.000 copies of an in vitro transcribed poly(A)-tailed RNA/cell). Analysis of total RNA and amplified cDNA, obtained from neutrophil granulocytes and the promyelocytic HL-60 cell line, demonst...Continue Reading

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Citations

Jan 28, 2004·Comptes rendus biologies·Jiangning LiRoger E Bumgarner
Apr 6, 2004·Progress in Neurobiology·David HinkleJames Eberwine
Jul 10, 2004·Molecular Plant-microbe Interactions : MPMI·Torben GjettingMichael F Lyngkjaer
Oct 29, 2005·BMC Genomics·Vigdis NygaardEivind Hovig
Jul 15, 2006·The Journal of Physiology·Kojiro YanoHugh P C Robinson
Sep 14, 2004·Leukemia & Lymphoma·Heidi Rye HudlebuschThomas Rasmussen
May 11, 2010·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Chiara GiudiceValeria Grieco
Jul 20, 2007·Proteomics·Qiangwei XiaMurray Hackett
Apr 4, 2015·The Journal of General Virology·Duan S LoyLyric C Bartholomay
Nov 30, 2005·Development·Masatake OsawaShin-Ichi Nishikawa
Aug 11, 2018·Molecular Therapy. Methods & Clinical Development·Aisha A AlJanahiCynthia E Dunbar
Mar 17, 2010·Biophysical Chemistry·Jens Christian BrasenLars Folke Olsen

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