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dc.contributor.authorRusso, Alessia
dc.contributor.authorMayjonade, Baptiste
dc.contributor.authorFrei, Daniel
dc.contributor.authorPotente, Giacomo
dc.contributor.authorKellenberger, Roman T
dc.contributor.authorFrachon, Léa
dc.contributor.authorCopetti, Dario
dc.contributor.authorStuder, Bruno
dc.contributor.authorFrey, Jürg E
dc.contributor.authorGrossniklaus, Ueli
dc.contributor.authorSchlüter, Philipp M
dc.date.accessioned2022-06-07T08:12:16Z
dc.date.available2022-06-07T08:12:16Z
dc.date.issued2022
dc.date.submitted2022-02-25
dc.identifier.issn1664-462X
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/337771
dc.description.abstractLong-read DNA sequencing technologies require high molecular weight (HMW) DNA of adequate purity and integrity, which can be difficult to isolate from plant material. Plant leaves usually contain high levels of carbohydrates and secondary metabolites that can impact DNA purity, affecting downstream applications. Several protocols and kits are available for HMW DNA extraction, but they usually require a high amount of input material and often lead to substantial DNA fragmentation, making sequencing suboptimal in terms of read length and data yield. We here describe a protocol for plant HMW DNA extraction from low input material (0.1 g) which is easy to follow and quick (2.5 h). This method successfully enabled us to extract HMW from four species from different families (Orchidaceae, Poaceae, Brassicaceae, Asteraceae). In the case of recalcitrant species, we show that an additional purification step is sufficient to deliver a clean DNA sample. We demonstrate the suitability of our protocol for long-read sequencing on the Oxford Nanopore Technologies PromethION® platform, with and without the use of a short fragment depletion kit.
dc.languageen
dc.publisherFrontiers Media SA
dc.subjectPlant Science
dc.subjectDNA extraction
dc.subjectDNA sequencing
dc.subjectnanopore sequencing
dc.subjectCirculomics
dc.subjectplant genome
dc.subjectONT long read sequencing
dc.subjectPacBio
dc.subjectgenome assembly
dc.titleLow-Input High-Molecular-Weight DNA Extraction for Long-Read Sequencing From Plants of Diverse Families.
dc.typeArticle
dc.date.updated2022-06-07T08:12:15Z
prism.publicationNameFront Plant Sci
prism.volume13
dc.identifier.doi10.17863/CAM.85180
dcterms.dateAccepted2022-04-21
rioxxterms.versionofrecord10.3389/fpls.2022.883897
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
dc.identifier.eissn1664-462X
cam.issuedOnline2022-05-19


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