Decoding DNA labels by melting curve analysis using real-time PCR

József A. Balog, Liliána Z. Fehér, L. Puskás

Research output: Contribution to journalArticle

Abstract

Synthetic DNA has been used as an authentication code for a diverse number of applications. However, existing decoding approaches are based on either DNA sequencing or the determination of DNA length variations. Here, we present a simple alternative protocol for labeling different objects using a small number of short DNA sequences that differ in their melting points. Code amplification and decoding can be done in two steps using quantitative PCR (qPCR). To obtain a DNA barcode with high complexity, we defined 8 template groups, each having 4 different DNA templates, yielding 158 (>2.5 billion) combinations of different individual melting temperature (Tm) values and corresponding ID codes. The reproducibility and specificity of the decoding was confirmed by using the most complex template mixture, which had 32 different products in 8 groups with different Tm values. The industrial applicability of our protocol was also demonstrated by labeling a drone with an oil-based paint containing a predefined DNA code, which was then successfully decoded. The method presented here consists of a simple code system based on a small number of synthetic DNA sequences and a cost-effective, rapid decoding protocol using a few qPCR reactions, enabling a wide range of authentication applications.

Original languageEnglish
Pages (from-to)261-266
Number of pages6
JournalBioTechniques
Volume63
Issue number6
DOIs
Publication statusPublished - Dec 1 2017

Fingerprint

Nucleic Acid Denaturation
Decoding
Labels
Real-Time Polymerase Chain Reaction
Melting
DNA
DNA sequences
Freezing
Labeling
Authentication
Melting point
Polymerase Chain Reaction
Temperature
Paint
Complex Mixtures
DNA Sequence Analysis
Oils
Amplification
Costs and Cost Analysis
Costs

Keywords

  • DNA coding
  • Melting curve analysis
  • Real-time PCR

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Decoding DNA labels by melting curve analysis using real-time PCR. / Balog, József A.; Fehér, Liliána Z.; Puskás, L.

In: BioTechniques, Vol. 63, No. 6, 01.12.2017, p. 261-266.

Research output: Contribution to journalArticle

Balog, József A. ; Fehér, Liliána Z. ; Puskás, L. / Decoding DNA labels by melting curve analysis using real-time PCR. In: BioTechniques. 2017 ; Vol. 63, No. 6. pp. 261-266.
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