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dc.contributor.authorStark, Jordan R.  Concept link
dc.contributor.authorCardon, Zoe G.  Concept link
dc.contributor.authorPeredo, Elena L.  Concept link
dc.date.accessioned2020-04-17T20:42:15Z
dc.date.available2020-04-17T20:42:15Z
dc.date.issued2020-03-10
dc.identifier.citationStark, J. R., Cardon, Z. G., & Peredo, E. L. (2020). Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae. Applications in Plant Sciences, 8(3), e11333.en_US
dc.identifier.urihttps://hdl.handle.net/1912/25656
dc.description© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Stark, J. R., Cardon, Z. G., & Peredo, E. L. Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae. Applications in Plant Sciences, 8(3), (2020): e11333, doi:10.1002/aps3.11333.en_US
dc.description.abstractPremise New sequencing technologies have facilitated genomic studies in green microalgae; however, extracting high‐quality DNA is often a bottleneck for long‐read sequencing. Methods and Results Here, we present a low‐cost, highly transferrable method for the extraction of high‐molecular‐weight (HMW), high‐purity DNA from microalgae. We first determined the effect of sample preparation on DNA quality using three homogenization methods: manual grinding using a mini‐pestle, automatic grinding using a vortex adapter, and grinding in liquid nitrogen. We demonstrated the versatility of grinding in liquid nitrogen followed by a modified cetyltrimethylammonium bromide (CTAB) extraction across a suite of aquatic‐ and desert‐evolved algal taxa. Finally, we tested the protocol's robustness by doubling the input material to increase yield, producing per sample up to 20 μg of high‐purity DNA longer than 21.2 kbp. Conclusions All homogenization methods produced DNA within acceptable parameters for purity, but only liquid nitrogen grinding resulted in HMW DNA. The optimization of cell lysis while minimizing DNA shearing is therefore crucial for the isolation of DNA for long‐read genomic sequencing because template DNA length strongly affects read output and length.en_US
dc.description.sponsorshipThe authors thank Dr. Louise Lewis (University of Connecticut) for providing Flechtneria rotunda and Acutodesmus deserticola, and Suzanne Thomas for expert technical assistance. This work was supported by the National Science Foundation, Division of Integrative Organismal Systems (1355085 to Z.G.C.) and an anonymous donor (to Z.G.C.).en_US
dc.publisherWiley Open Accessen_US
dc.relation.urihttps://doi.org/10.1002/aps3.11333
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectDNA integrityen_US
dc.subjectlong‐read sequencingen_US
dc.subjectmodified CTAB extractionen_US
dc.subjectScenedesmaceaeen_US
dc.titleExtraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgaeen_US
dc.typeArticleen_US
dc.identifier.doi10.1002/aps3.11333


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Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivatives 4.0 International