Document Detail


Genome-wide transcriptome analysis of 150 cell samples.
MedLine Citation:
PMID:  20023796     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A major challenge in molecular biology is interrogating the human transcriptome on a genome wide scale when only a limited amount of biological sample is available for analysis. Current methodologies using microarray technologies for simultaneously monitoring mRNA transcription levels require nanogram amounts of total RNA. To overcome the sample size limitation of current technologies, we have developed a method to probe the global gene expression in biological samples as small as 150 cells, or the equivalent of approximately 300 pg total RNA. The new method employs microfluidic devices for the purification of total RNA from mammalian cells and ultra-sensitive whole transcriptome amplification techniques. We verified that the RNA integrity is preserved through the isolation process, accomplished highly reproducible whole transcriptome analysis, and established high correlation between repeated isolations of 150 cells and the same cell culture sample. We validated the technology by demonstrating that the combined microfluidic and amplification protocol is capable of identifying biological pathways perturbed by stimulation, which are consistent with the information recognized in bulk-isolated samples.
Authors:
Daniel Irimia; Michael Mindrinos; Aman Russom; Wenzhong Xiao; Julie Wilhelmy; Shenglong Wang; Joe Don Heath; Nurith Kurn; Ronald G Tompkins; Ronald W Davis; Mehmet Toner
Related Documents :
10069996 - Polyamine depletion arrests cell cycle and induces inhibitors p21(waf1/cip1), p27(kip1)...
12450656 - Analysis of polyamines as markers of (patho)physiological conditions.
1346156 - Defective rnas in mosquito cells persistently infected with bunyamwera virus.
7689156 - Depletion of cellular polyamines by alpha-difluoromethylornithine increases 1,3-bis(2-c...
7279786 - Scanning electron microscopy of normal, malignant and post radiotherapeutic oral mucosa...
7643716 - Effects of doxorubicin on cardiomyocytes with reduced level of superoxide dismutase.
Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, N.I.H., Extramural     Date:  2008-11-12
Journal Detail:
Title:  Integrative biology : quantitative biosciences from nano to macro     Volume:  1     ISSN:  1757-9708     ISO Abbreviation:  Integr Biol (Camb)     Publication Date:  2009 Jan 
Date Detail:
Created Date:  2009-12-21     Completed Date:  2010-03-10     Revised Date:  2013-09-10    
Medline Journal Info:
Nlm Unique ID:  101478378     Medline TA:  Integr Biol (Camb)     Country:  England    
Other Details:
Languages:  eng     Pagination:  99-107     Citation Subset:  IM    
Affiliation:
BioMEMS Resource Center, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, 02114, USA. dirimia@hms.harvard.edu
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Cell Separation / instrumentation*,  methods
Chromosome Mapping / instrumentation*,  methods
Equipment Design
Equipment Failure Analysis
Gene Expression Profiling / instrumentation*,  methods
Microfluidic Analytical Techniques / instrumentation*,  methods
Nucleic Acid Amplification Techniques / instrumentation*,  methods
RNA / genetics*,  isolation & purification
Grant Support
ID/Acronym/Agency:
P01 HG000205/HG/NHGRI NIH HHS; P01HG000205/HG/NHGRI NIH HHS; P41 EB002503/EB/NIBIB NIH HHS; U54 GM062119/GM/NIGMS NIH HHS; U54GM62119/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
63231-63-0/RNA
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Real-time detection of the early event of cytotoxicity of herbal ingredients on single leukemia cell...
Next Document:  Influence of applied currents on the viability of cells close to microelectrodes.