Document Detail


Fiber-optic DNA sensor for fluorometric nucleic acid determination.
MedLine Citation:
PMID:  8849028     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Single-stranded deoxyribonucleic acid (ssDNA) thymidylic acid icosanucleotides (dT20) were synthesized on the surfaces of derivatized quartz optical fibers to create an optical DNA biosensor. The synthesis made use of an automated solid-phase synthesizer and phosphoramidite synthons. The covalently immobilized oligomers were found to hybridize with complementary ssDNA (cDNA) or ssRNA (cRNA) from solution, and the device was regenerable for multiple cycles of application. Hybridization on optical fibers was detected by the use of the fluorescent DNA stain ethidium bromide (EB). The procedure used hybridization assay techniques and provided a detection limit of 86 ng x mL(-1) cDNA and a sensitivity of 200% fluorescence intensity increase per 100 ng x mL(-1) of cDNA, with one cycle of hybridization analysis requiring 45 min. The sensor has been observed to be regenerable (minimum of five cycles) and to sustain full activity after prolonged storage times (1 year), harsh washing conditions (sonication), and sterilization (autoclaving). The extent of hybridization between the immobilized and complementary nucleic acid strands was determined by UV absorbance thermal denaturation studies wherein all 20 bases on each strand of the nucleic acid were found to be involved in duplex formation.
Authors:
P A Piunno; U J Krull; R H Hudson; M J Damha; H Cohen
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  67     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  1995 Aug 
Date Detail:
Created Date:  1996-10-21     Completed Date:  1996-10-21     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2635-43     Citation Subset:  IM    
Affiliation:
Chemical Sensors Group, Department of Chemistry, Erindale College, Univerity of Toronto, Ontario, Canada.
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MeSH Terms
Descriptor/Qualifier:
Biosensing Techniques
DNA / analysis*
Fiber Optic Technology
Nucleic Acid Hybridization
Oligonucleotides / chemical synthesis,  chemistry
Optical Fibers
Spectrometry, Fluorescence
Chemical
Reg. No./Substance:
0/Oligonucleotides; 9007-49-2/DNA

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


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