DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen
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
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BiotechniquesDate
2020Copyright
© Authors, 2020
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.
Publisher
Future Science LtdISSN Search the Publication Forum
0736-6205Keywords
Publication in research information system
https://converis.jyu.fi/converis/portal/detail/Publication/43379751
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Related funder(s)
Research Council of Finland; European CommissionFunding program(s)
Academy Project, AoF; FP7 (EU's 7th Framework Programme); ERC Proof of Concept Grant
The content of the publication reflects only the author’s view. The funder is not responsible for any use that may be made of the information it contains.
Additional information about funding
This work was funded by the Academy of Finland (grant 323063) and European Research Council CoG grant (615146) and PoC grant (812729; EU FP7 and H2020 programs, respectively) to M Tiirola.License
Except where otherwise noted, this item's license is described as CC BY 4.0