Document Detail


BeadCons: detection of nucleic acid sequences by flow cytometry.
MedLine Citation:
PMID:  18770821     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Molecular beacons are single-stranded nucleic acid structures with a terminal fluorophore and a distal, terminal quencher. These molecules are typically used in real-time PCR assays, but have also been conjugated with solid matrices. This unit describes protocols related to molecular beacon-conjugated beads (BeadCons), whose specific hybridization with complementary target sequences can be resolved by cytometry. Assay sensitivity is achieved through the concentration of fluorescence signal on discrete particles. By using molecular beacons with different fluorophores and microspheres of different sizes, it is possible to construct a fluid array system with each bead corresponding to a specific target nucleic acid. Methods are presented for the design, construction, and use of BeadCons for the specific, multiplexed detection of unlabeled nucleic acids in solution. The use of bead-based detection methods will likely lead to the design of new multiplex molecular diagnostic tools.
Authors:
Douglas Horejsh; Federico Martini; Maria Rosaria Capobianchi
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Current protocols in cytometry / editorial board, J. Paul Robinson, managing editor ... [et al.]     Volume:  Chapter 13     ISSN:  1934-9300     ISO Abbreviation:  Curr Protoc Cytom     Publication Date:  2005 Nov 
Date Detail:
Created Date:  2008-09-04     Completed Date:  2008-10-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100899351     Medline TA:  Curr Protoc Cytom     Country:  United States    
Other Details:
Languages:  eng     Pagination:  Unit13.5     Citation Subset:  IM    
Affiliation:
National Institute for Infectious Diseases "L. Spallazani"-IRCSS, Rome, Italy.
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MeSH Terms
Descriptor/Qualifier:
Base Sequence*
DNA / chemistry*,  genetics
Flow Cytometry / methods*
Fluorescent Dyes
Nucleic Acid Conformation
Nucleic Acid Hybridization
Oligonucleotide Array Sequence Analysis / methods
Oligonucleotide Probes
Sensitivity and Specificity
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 0/Oligonucleotide Probes; 9007-49-2/DNA

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


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