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JoVE Journal

Genetics

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

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12:33 min

July 28th, 2017

July 28th, 2017

12,765 Views

0:05

Title

0:49

Growing Wheat Plants

1:51

Organellar DNA Enrichment by Differential Centrifugation (DC)

7:31

Organellar DNA Enrichment by Methyl-fractionation (MF)

9:44

Results: Both Enrichment Methods Yield Sufficient DNA Highly Enriched for Organellar Sequences and Suitable for NGS

11:33

Conclusion

Transcript

The overall goal of this experiment is to compare two methods used to isolate high quality organellar DNA, both plastid and mitochondria DNA from plant leaf tissue that is suitable for next-generation sequencing. These methods can help answer key

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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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