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  TaqMan probe array for quantitative detection of DNA targets

Liu, H., Wang, H., Shi, Z., Wang, H., Yang, C., Sperling, S., et al. (2006). TaqMan probe array for quantitative detection of DNA targets. Nucleic Acids Research (London), 34, e4-e4. doi:10.1093/nar/gnj006.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-84CF-1 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-84D0-C
Genre: Journal Article
Alternative Title : Nucleic Acids Res

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 Creators:
Liu, Heping, Author
Wang, Hong, Author
Shi, Zhiyang, Author
Wang, Hua, Author
Yang, Chaoyong, Author
Sperling, Silke1, Author              
Tan, Weihong, Author
Lu, Zuhong, Author
Affiliations:
1Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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 Abstract: To date real-time quantitative PCR and gene expression microarrays are the methods of choice for quantification of nucleic acids. Herein, we described a unique fluorescence resonance energy transfer-based microarray platform for real-time quantification of nucleic acid targets that combines advantages of both and reduces their limitations. A set of 3' amino-modified TaqMan probes were designed and immobilized on a glass slide composing a regular microarray pattern, and used as probes in the consecutive PCR carried out on the surface. During the extension step of the PCR, 5' nuclease activity of DNA polymerase will cleave quencher dyes of the immobilized probe in the presence of nucleic acids targets. The increase of fluorescence intensities generated by the change in physical distance between reporter fluorophore and quencher moiety of the probes were collected by a confocal scanner. Using this new approach we successfully monitored five different pathogenic genomic DNAs and analyzed the dynamic characteristics of fluorescence intensity changes on the TaqMan probe array. The results indicate that the TaqMan probe array on a planar glass slide monitors DNA targets with excellent specificity as well as high sensitivity. This set-up offers the great advantage of real-time quantitative detection of DNA targets in a parallel array format.

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Language(s): eng - English
 Dates: 2006-01-10
 Publication Status: Published in print
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 Table of Contents: -
 Rev. Type: -
 Identifiers: eDoc: 309273
DOI: 10.1093/nar/gnj006
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Title: Nucleic Acids Research (London)
  Alternative Title : Nucleic Acids Res
Source Genre: Journal
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Pages: - Volume / Issue: 34 Sequence Number: - Start / End Page: e4 - e4 Identifier: ISSN: 1362-4962