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Heat transfer time determination based on DNA melting curve analysis

Abstract

The determination of the physical properties of fluids—such as the thermal characteristics, which include heat transfer time (Δt)—is becoming more challenging as system sizes shrink to micro- and nanometer scales. Hence, knowledge of these properties is crucial for the operation of devices requiring precise temperature (T) control, such as polymerase chain reactions, melting curve analysis (MCA), and differential scanning fluorimetry. In this paper, we introduced a flow-through microfluidic system to analyze thermal properties such as Δt among samples and the sidewall of a silicon chip using microscopic image analysis. We performed a spatial MCA with double-stranded deoxyribonucleic acid (dsDNA) and EvaGreen intercalator, using a flow-through microfluidic chip, and achieved a T gradient of ≈ 2.23 K mm−1. We calculated the mean value of Δt as ≈ 33.9 ms from a melting temperature (TM) location shift along the microchannel for a variable flow rate. Our system had a T resolution of ≈ 1.2 mK pixel−1 to distinguish different dsDNA molecules—based on the TM location within the chip—providing an option to use it as a high-throughput device for rapid DNA or protein analysis.

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Acknowledgements

The authors wish to acknowledge the financial support of the W099109 grant from P.R. China, the GACR Project Number GA16-11140S from the Czech Republic and the technical support provided by Nano+ and by Centrum SIX of Brno University of Technology, Czech Republic. LY and SN acknowledge the funding from the Research Grant Council of Hong Kong, received under Grant Number 16209316.

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Correspondence to Pavel Neuzil.

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Zhu, H., Li, H., Zhang, H. et al. Heat transfer time determination based on DNA melting curve analysis. Microfluid Nanofluid 24, 7 (2020). https://doi.org/10.1007/s10404-019-2308-9

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  • DOI: https://doi.org/10.1007/s10404-019-2308-9

Keywords

  • dsDNA
  • Melting curve analysis
  • Heat transfer
  • Microfluidics