A1 Journal article (refereed)
DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen (2020)

Dutra, L., Franz, O., Puupponen, V.-M., & Tiirola, M. (2020). DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen. Biotechniques, 69(9), 451-454. https://doi.org/10.2144/btn-2020-0076

JYU authors or editors

Ambrosio Leal Dutra, Lara
Franz, Ole
Puupponen, Veli-Mikko
Tiirola, Marja

Publication details

All authors or editors: Dutra, Lara; Franz, Ole; Puupponen, Veli-Mikko; Tiirola, Marja

Journal or series: Biotechniques

ISSN: 0736-6205

eISSN: 1940-9818

Publication year: 2020

Publication date: 26/10/2020

Volume: 69

Issue number: 9

Pages range: 451-454

Publisher: Future Science Ltd

Publication country: United Kingdom

Publication language: English

DOI: https://doi.org/10.2144/btn-2020-0076

Publication open access: Openly available

Publication channel open access: Open Access channel

Publication is parallel published (JYX): https://jyx.jyu.fi/handle/123456789/72402

Abstract

Droplet microfluidics is a technology that enables the production and manipulation of small volumes. In biosciences, the most popular application of this technology is Droplet Digital™ PCR (ddPCR™), where parallel nanoliter-scale PCR assays are used to provide a high sensitivity and specificity for DNA detection. However, the recovery of PCR products for downstream applications such as sequencing can be challenging due to the droplets' stability. Here we compared five methods for disrupting the droplets to recover DNA. We found that rapid freezing in liquid nitrogen results in a clear phase separation and recovery of up to 70% of the DNA content. Liquid nitrogen freezing can thus offer a simple and environmentally friendly protocol for recovering DNA from ddPCR.

Keywords: biotechnology; laboratory technology; microfluidics; DNA

Free keywords: amplicon recovery; breaking droplets; Droplet Digital™ PCR (ddPCR™); droplet microfluidics; emulsion PCR

Fields of science: