Protein detection via direct enzymatic amplification of short DNA aptamers

Nicholas O Fischer, Theodore M. Tarasow, Jeffrey B H Tok

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

Aptamers are single-stranded nucleic acids that fold into defined tertiary structures to bind target molecules with high specificities and affinities. DNA aptamers have garnered much interest as recognition elements for biodetection and diagnostic applications due to their small size, ease of discovery and synthesis, and chemical and thermal stability. Here we describe the design and application of a short DNA molecule capable of both protein target binding and amplifiable bioreadout processes. Because both recognition and readout capabilities are incorporated into a single DNA molecule, tedious conjugation procedures required for protein-DNA hybrids can be omitted. The DNA aptamer is designed to be amplified directly by either polymerase chain reaction (PCR) or rolling circle amplification (RCA) processes, taking advantage of real-time amplification monitoring techniques for target detection. A combination of both RCA and PCR provides a wide protein target dynamic range (1 μM to 10 pM).

Original languageEnglish (US)
Pages (from-to)121-128
Number of pages8
JournalAnalytical Biochemistry
Volume373
Issue number1
DOIs
StatePublished - Feb 1 2008
Externally publishedYes

Fingerprint

Nucleotide Aptamers
Amplification
Polymerase chain reaction
Molecules
DNA
Polymerase Chain Reaction
Proteins
Chemical stability
Target tracking
Protein Binding
Nucleic Acids
Thermodynamic stability
Hot Temperature
Monitoring

Keywords

  • Aptamers
  • Limit of detection
  • PCR
  • Rolling circle amplification

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Protein detection via direct enzymatic amplification of short DNA aptamers. / Fischer, Nicholas O; Tarasow, Theodore M.; Tok, Jeffrey B H.

In: Analytical Biochemistry, Vol. 373, No. 1, 01.02.2008, p. 121-128.

Research output: Contribution to journalArticle

Fischer, Nicholas O ; Tarasow, Theodore M. ; Tok, Jeffrey B H. / Protein detection via direct enzymatic amplification of short DNA aptamers. In: Analytical Biochemistry. 2008 ; Vol. 373, No. 1. pp. 121-128.
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