DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen

Dutra, Lara; Franz, Ole; Puupponen, Veli-Mikko; Tiirola, Marja

doi:10.2144/btn-2020-0076

dc.contributor.author	Dutra, Lara
dc.contributor.author	Franz, Ole
dc.contributor.author	Puupponen, Veli-Mikko
dc.contributor.author	Tiirola, Marja
dc.date.accessioned	2020-10-30T09:59:00Z
dc.date.available	2020-10-30T09:59:00Z
dc.date.issued	2020
dc.identifier.citation	Dutra, L., Franz, O., Puupponen, V.-M., & Tiirola, M. (2020). DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen. <i>Biotechniques</i>, <i>69</i>(9), 451-454. <a href="https://doi.org/10.2144/btn-2020-0076" target="_blank">https://doi.org/10.2144/btn-2020-0076</a>
dc.identifier.other	CONVID_43379751
dc.identifier.uri	https://jyx.jyu.fi/handle/123456789/72402
dc.description.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.	en
dc.format.mimetype	application/pdf
dc.language	eng
dc.language.iso	eng
dc.publisher	Future Science Ltd
dc.relation.ispartofseries	Biotechniques
dc.rights	CC BY 4.0
dc.subject.other	amplicon recovery
dc.subject.other	breaking droplets
dc.subject.other	Droplet Digital™ PCR (ddPCR™)
dc.subject.other	droplet microfluidics
dc.subject.other	emulsion PCR
dc.title	DNA recovery from Droplet Digital™ PCR emulsions using liquid nitrogen
dc.type	article
dc.identifier.urn	URN:NBN:fi:jyu-202010306447
dc.contributor.laitos	Bio- ja ympäristötieteiden laitos	fi
dc.contributor.laitos	Department of Biological and Environmental Science	en
dc.contributor.oppiaine	Ympäristötiede	fi
dc.contributor.oppiaine	Solu- ja molekyylibiologia	fi
dc.contributor.oppiaine	Environmental Science	en
dc.contributor.oppiaine	Cell and Molecular Biology	en
dc.type.uri	http://purl.org/eprint/type/JournalArticle
dc.type.coar	http://purl.org/coar/resource_type/c_2df8fbb1
dc.description.reviewstatus	peerReviewed
dc.format.pagerange	451-454
dc.relation.issn	0736-6205
dc.relation.numberinseries	9
dc.relation.volume	69
dc.type.version	publishedVersion
dc.rights.copyright	© Authors, 2020
dc.rights.accesslevel	openAccess	fi
dc.relation.grantnumber	323063
dc.relation.grantnumber	615146
dc.relation.grantnumber	615146
dc.relation.grantnumber	812729
dc.relation.grantnumber	812729
dc.relation.projectid	info:eu-repo/grantAgreement/EC/FP7/615146/EU//
dc.relation.projectid	info:eu-repo/grantAgreement/EC/H2020/812729/EU//BiopSense
dc.subject.yso	mikrofluidistiikka
dc.subject.yso	biotekniikka
dc.subject.yso	laboratoriotekniikka
dc.subject.yso	DNA
dc.format.content	fulltext
jyx.subject.uri	http://www.yso.fi/onto/yso/p38414
jyx.subject.uri	http://www.yso.fi/onto/yso/p2348
jyx.subject.uri	http://www.yso.fi/onto/yso/p19594
jyx.subject.uri	http://www.yso.fi/onto/yso/p7690
dc.rights.url	https://creativecommons.org/licenses/by/4.0/
dc.relation.doi	10.2144/btn-2020-0076
dc.relation.funder	Research Council of Finland	en
dc.relation.funder	European Commission	en
dc.relation.funder	European Commission	en
dc.relation.funder	Suomen Akatemia	fi
dc.relation.funder	Euroopan komissio	fi
dc.relation.funder	Euroopan komissio	fi
jyx.fundingprogram	Academy Project, AoF	en
jyx.fundingprogram	FP7 (EU's 7th Framework Programme)	en
jyx.fundingprogram	ERC Proof of Concept Grant	en
jyx.fundingprogram	Akatemiahanke, SA	fi
jyx.fundingprogram	EU:n 7. puiteohjelma (FP7)	fi
jyx.fundingprogram	ERC Proof of Concept Grant	fi
jyx.fundinginformation	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.
dc.type.okm	A1