Journal
/
Genetics
/
Optimization and Comparative Analysis of Plant Organellar DNA Enrichment Methods Suitable for Next-generation Sequencing
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
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

DOI:
10.3791/55528-v

•

12:33 min

•

July 28, 2017

•

, ,
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

Chapters

  • 00:05Title
  • 00:49Growing Wheat Plants
  • 01:51Organellar DNA Enrichment by Differential Centrifugation (DC)
  • 07:31Organellar DNA Enrichment by Methyl-fractionation (MF)
  • 09:44Results: Both Enrichment Methods Yield Sufficient DNA Highly Enriched for Organellar Sequences and Suitable for NGS
  • 11:33Conclusion

Summary

Automatic Translation

Automatic Translation

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.

Tags

Plant OrganelleDNA EnrichmentNext-generation SequencingDifferential CentrifugationMeta FractionationWheat SeedlingOrganelle IsolationSTE BufferFiltration

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

Methyl-binding DNA capture Sequencing for Patient Tissues
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

Assessment of DNA Contamination in RNA Samples Based on Ribosomal DNA
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

Robust DNA Isolation and High-throughput Sequencing Library Construction for Herbarium Specimens
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

Rare Event Detection Using Error-corrected DNA and RNA Sequencing
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

Amplification of Near Full-length HIV-1 Proviruses for Next-Generation Sequencing
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq)
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

RNA Next-Generation Sequencing and a Bioinformatics Pipeline to Identify Expressed LINE-1s at the Locus-Specific Level
check_url/55528?article_type=v&slug=optimization-comparative-analysis-plant-organellar-dna-enrichment

Single-Molecule Förster Resonance Energy Transfer Methods for Real-Time Investigation of the Holliday Junction Resolution by GEN1

Read Article