Document Detail


mRNA Quantification After Fluorescence Activated Cell Sorting Using Locked Nucleic Acid Probes.
MedLine Citation:
PMID:  21246309     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Recently, we have established an in-tube in situ hybridization method named mRNA quantification after fluorescence activated cell sorting (FACS-mQ), in which a specific RNA in a particular cell type is stained with a florescent dye, allowing the stained cells to be selected by FACS without suffering excessive RNA degradation. Using this method, the biological characteristics of the sorted cells can be determined by analyzing their gene expression profile. In this study, we used locked nucleic acid (LNA) oligonucleotides, which are known to enhance both the sensitivity and specificity of RNA detection, as hybridization probes in FACS-mQ. When we used a LNA probe targeting the human 28S sequence, we were able to efficiently separate human cells from rat cells. Using LNA probes, the hybridization step was shortened to 1 h. After the hybridization step, 84.6% RNA was preserved; thus, we were able to successfully measure gene expression levels in each type of cell after FACS. Providing the LNA probe efficiently hybridizes with the target sequence, FACS-mQ with an LNA probe is a powerful tool for separating particular cells and determining their biological characteristics by analyzing their gene expression profile.
Authors:
Rie Maruo; Hiroya Yamada; Mikio Watanabe; Yoh Hidaka; Yoshinori Iwatani; Toru Takano
Related Documents :
21262849 - Ixabepilone, a novel microtubule-targeting agent for breast cancer, is a substrate for ...
25228569 - Developmental anatomy and immunocytochemistry reveal the neo-ontogenesis of the leaf ti...
25171109 - Reduction of cardiac cell death after helium postconditioning in rats: transcriptional ...
25142939 - Engager t cells: a new class of antigen-specific t cells that redirect bystander t cells.
25227819 - Cytotoxicity of trichoderma spp. cultural filtrate against human cervical and breast ca...
25450809 - Interferon-γ induces a tryptophan-selective amino acid transporter in human colonic ep...
18197149 - Second hit post burn increased proximal gut mucosa epithelial cells damage.
16601139 - Chronic palmitate but not oleate exposure induces endoplasmic reticulum stress, which m...
2612369 - Re-expression of 117 antigen, a cell surface glycoprotein of aggregating cells, during ...
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-1-19
Journal Detail:
Title:  Molecular biotechnology     Volume:  -     ISSN:  1559-0305     ISO Abbreviation:  -     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2011-1-19     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9423533     Medline TA:  Mol Biotechnol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Department of Laboratory Medicine, Osaka University Graduate School of Medicine, D2, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan, marucchi_stone0611@yahoo.co.jp.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:

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


Previous Document:  The Etiology of Paraneoplastic Autoimmunity.
Next Document:  Targeted IV busulfan and fludarabine followed by post-allogeneic hematopoietic cell transplantation ...