DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen
| Autor: | Lara Dutra, Ole Franz, Veli-Mikko Puupponen, Marja Tiirola |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 69, Iss 6, Pp 450-454 (2020) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 03261581 |
| DOI: | 10.2144/btn-2020-0076 |
| Popis: | 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. |
| Databáze: | Directory of Open Access Journals |
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