Enzymatically amplified SPR imaging detection of DNA by exonuclease III digestion of DNA microarrays

H.J. Lee, Y. Li, Alastair Wark, R.M. Corn

Research output: Contribution to journalArticle

144 Citations (Scopus)

Abstract

This paper describes a novel approach utilizing the enzyme exonuclease III in conjunction with 3‘-terminated DNA microarrays for the amplified detection of single-stranded DNA (ssDNA) with surface plasmon resonance (SPR) imaging. When ExoIII and target DNA are simultaneously introduced to a 3‘-terminated ssDNA microarray, hybridization adsorption of the target ssDNA leads to the direction-dependent ExoIII hydrolysis of probe ssDNA strands and the release of the intact target ssDNA back into the solution. Readsorption of the target ssDNA to another probe creates a repeated hydrolysis process that results over time in a significant negative change in SPR imaging signal. Experiments are presented that demonstrate the direction-dependent surface enzyme reaction of ExoIII with double-stranded DNA as well as this new enzymatically amplified SPR imaging process with a 16-mer target ssDNA detection limit of 10−100 pM. This is a 102−103 improvement on previously reported measurements of SPR imaging detection of ssDNA based solely on hybridization adsorption without enzymatic amplification
LanguageEnglish
Pages5096-5100
Number of pages5
JournalAnalytical Chemistry
Volume77
Issue number16
Early online date30 Jun 2005
DOIs
Publication statusPublished - 2005

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Exodeoxyribonucleases
Surface Plasmon Resonance
Single-Stranded DNA
Surface plasmon resonance
Microarrays
Oligonucleotide Array Sequence Analysis
Digestion
Imaging techniques
DNA
Adsorption
Hydrolysis
exodeoxyribonuclease III
Enzymes
Amplification
Limit of Detection

Keywords

  • exonuclease
  • enzymatically amplified
  • SPR imaging detection
  • DNA
  • digestion
  • dna microarrays

Cite this

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title = "Enzymatically amplified SPR imaging detection of DNA by exonuclease III digestion of DNA microarrays",
abstract = "This paper describes a novel approach utilizing the enzyme exonuclease III in conjunction with 3‘-terminated DNA microarrays for the amplified detection of single-stranded DNA (ssDNA) with surface plasmon resonance (SPR) imaging. When ExoIII and target DNA are simultaneously introduced to a 3‘-terminated ssDNA microarray, hybridization adsorption of the target ssDNA leads to the direction-dependent ExoIII hydrolysis of probe ssDNA strands and the release of the intact target ssDNA back into the solution. Readsorption of the target ssDNA to another probe creates a repeated hydrolysis process that results over time in a significant negative change in SPR imaging signal. Experiments are presented that demonstrate the direction-dependent surface enzyme reaction of ExoIII with double-stranded DNA as well as this new enzymatically amplified SPR imaging process with a 16-mer target ssDNA detection limit of 10−100 pM. This is a 102−103 improvement on previously reported measurements of SPR imaging detection of ssDNA based solely on hybridization adsorption without enzymatic amplification",
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Enzymatically amplified SPR imaging detection of DNA by exonuclease III digestion of DNA microarrays. / Lee, H.J.; Li, Y.; Wark, Alastair; Corn, R.M.

In: Analytical Chemistry, Vol. 77, No. 16, 2005, p. 5096-5100.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Enzymatically amplified SPR imaging detection of DNA by exonuclease III digestion of DNA microarrays

AU - Lee, H.J.

AU - Li, Y.

AU - Wark, Alastair

AU - Corn, R.M.

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AB - This paper describes a novel approach utilizing the enzyme exonuclease III in conjunction with 3‘-terminated DNA microarrays for the amplified detection of single-stranded DNA (ssDNA) with surface plasmon resonance (SPR) imaging. When ExoIII and target DNA are simultaneously introduced to a 3‘-terminated ssDNA microarray, hybridization adsorption of the target ssDNA leads to the direction-dependent ExoIII hydrolysis of probe ssDNA strands and the release of the intact target ssDNA back into the solution. Readsorption of the target ssDNA to another probe creates a repeated hydrolysis process that results over time in a significant negative change in SPR imaging signal. Experiments are presented that demonstrate the direction-dependent surface enzyme reaction of ExoIII with double-stranded DNA as well as this new enzymatically amplified SPR imaging process with a 16-mer target ssDNA detection limit of 10−100 pM. This is a 102−103 improvement on previously reported measurements of SPR imaging detection of ssDNA based solely on hybridization adsorption without enzymatic amplification

KW - exonuclease

KW - enzymatically amplified

KW - SPR imaging detection

KW - DNA

KW - digestion

KW - dna microarrays

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JO - Analytical Chemistry

T2 - Analytical Chemistry

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