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Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing
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Genetics
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JoVE Journal Genetics
Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing

Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing

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12,723 Views

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12:33 min

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July 28, 2017

DOI:

10.3791/55528-v

DOI:
10.3791/55528-v

•

12:33 min
July 28, 2017

•

16 Views
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1Cereal Disease Laboratory,United States Department of Agriculture-Agricultural Research Service, 2Department of Horticultural Science,University of Minnesota, 3Department of Plant Pathology,University of Minnesota

Summary

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The comparison and optimization of two plant organellar DNA enrichment methods are presented: traditional differential centrifugation and fractionation of the total gDNA based on methylation status. We assess the resulting DNA quantity and quality, demonstrate performance in short-read next-generation sequencing, and discuss the potential for use in long-read single-molecule sequencing.

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Miller, M. E., Liberatore, K. L., Kianian, S. F. Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing. J. Vis. Exp. (125), e55528, doi:10.3791/55528 (2017).

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