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An integrated droplet digital PCR gene chip for absolute quantification of nucleic acid

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Abstract

An integrated droplet digital polymerase chain reaction (ddPCR) gene chip that can quantify nucleic acid absolutely was constructed to avoid problems such as droplet fusion, fragmentation and sample contamination during pipetting. The integrated ddPCR gene chip was fabricated by photolithography and cyclic olefin copolymer (COC) injection mold. The 20 μL sample was distributed to about 60,000 water-in-oil droplets with a diameter of about 87 μm within 3 min by flow focus microstructure, and the volume of each droplet was about 0.27 nL. The generation of bubbles was reduced by reducing the width of the collection chamber and increasing the deceleration step. In addition, the efficiency of PCR was improved using COC materials. The sample of human epidermal growth factor receptor (EGFR) exon18 gene was quantitatively detected by this chip. Experimental results show that the detection signal has a good linear relationship with the DNA concentration from 101 to 105 copies/μL (R2 = 0.999) so that the integrated ddPCR gene chip can realize absolute quantification of nucleic acid. It is verified that the integration of droplet generation, PCR amplification and fluorescence detection are realized on one chip, reduce bubble generation and improve PCR efficiency. Compared with the current independent split chip, the integrated ddPCR gene chip ensures a high degree of anti-pollution performance, avoids the operation of droplet movement, reduces human interference, and standardizes the results. It has wide application prospects in nucleic acid quantification and trace detection.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work described in this paper was supported by the Jilin Scientific and Technological Development Program (CN) (Grant Number 20191102003YY).

Funding

This study was funded by the Jilin Scientific and Technological Development Program (CN) (Grant Number 20191102003YY).

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Authors and Affiliations

  1. Changchun University of Science and Technology, Changchun, 130022, China

    Xiangkai Meng, Yuanhua Yu, Ping Gong & Guangyong Jin

  2. School of Science, Changchun University of Science and Technology, Changchun, 130022, China

    Xiangkai Meng & Guangyong Jin

  3. Key Laboratory of Biological Detection Engineering, Changchun, 130022, China

    Yuanhua Yu & Ping Gong

Authors
  1. Xiangkai Meng
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  2. Yuanhua Yu
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  3. Ping Gong
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  4. Guangyong Jin
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Contributions

GJ contributed to the conception of the study, YY and PG performed the experiment, XM contributed data analysis and wrote the manuscript.

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Correspondence to Guangyong Jin.

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Meng, X., Yu, Y., Gong, P. et al. An integrated droplet digital PCR gene chip for absolute quantification of nucleic acid. Microfluid Nanofluid 25, 62 (2021). https://doi.org/10.1007/s10404-021-02465-4

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  • DOI: https://doi.org/10.1007/s10404-021-02465-4

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