ArticlePlant methods2020
A simple plant high-molecular-weight DNA extraction method suitable for single-molecule technologies.
Article in Plant methods, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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17 citing papers in PubMed, 48 citations in OpenAlex.
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- A high-quality genome assembly of the tetraploid Teucrium chamaedrys unveils a recent whole-genome duplication and a large biosynthetic gene cluster for diterpenoid metabolism.Plant communications · 2025Article
- The addition of Psathyrostachys Huashanica Keng 6Ns large segment chromosomes has positive impact on stripe rust resistance and plant spikelet number of common wheat.BMC plant biology · 2024Article
- Genome resources for three modern cotton lines guide future breeding efforts.Nature plants · 2024Article
- Sequence Characterization of Extra-Chromosomal Circular DNA Content in Multiple Blackgrass (Genes · 2023Article
- Development of High-Quality Nuclei Isolation to Study Plant Root-Microbe Interaction for Single-Nuclei Transcriptomic Sequencing in Soybean.Plants (Basel, Switzerland) · 2023Article
- A high quality, high molecular weight DNA extraction method for PacBio HiFi genome sequencing of recalcitrant plants.Plant methods · 2023Article
- Diversity of the repetitive DNA fraction in Cestrum, the genus with the largest genomes within Solanaceae.Molecular biology reports · 2022Article
- Less is more: natural variation disrupting a miR172 gene at the di locus underlies the recessive double-flower trait in peach (P. persica L. Batsch).BMC plant biology · 2022Article
- Plant genome sequence assembly in the era of long reads: Progress, challenges and future directions.Quantitative plant biology · 2022Review
- Rapid identification and quantitation of single plant seed allergen using paper-based microfluidics.PloS one · 2022Article
- Highly accurate long-read HiFi sequencing data for five complex genomes.Scientific data · 2020Article
- High-molecular-weight DNA extraction for long-read sequencing of plant genomes: An optimization of standard methods.Applications in plant sciencesArticle
- Modified CTAB protocols for high-molecular-weight DNA extractions from ferns.Applications in plant sciencesArticle
- Balancing read length and sequencing depth: Optimizing Nanopore long-read sequencing for monocots with an emphasis on the Liliales.Applications in plant sciencesArticle
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundHigh-molecular-weight and pure DNA is crucial for high-quality results from 3rd generation DNA Analyzers and optical mapping technologies. Conventional nuclei isolation methods for preparing high-molecular-weight genomic DNA from plant tissues include the preparation of protoplasts or embedding nuclei in an agarose matrix with subsequent manipulations via electro-elution or pulsed-field gel electrophoresis.
resultsIn this method, plant nuclei are isolated by physically grinding tissues and reconstituting the intact nuclei in a unique Nuclear Isolation Buffer (NIB). The plastid DNAs are released from organelles and eliminated with an osmotic buffer by washing and centrifugation. The purified nuclei are then lysed and further cleaned by organic extraction, and the genomic DNA is precipitated with a high concentration of CTAB. The highly pure, high molecular weight gDNA is extracted from the nuclei, dissolved in a high pH buffer, allowing for stable long-term storage.
conclusionsThis method is unique and avoids the use of embedding in agarose, which dramatically reduces time (4-8 h versus days), complexity, and materials cost. This procedure can be used on essentially any plant species and tissue stage. Here we describe a case study and a simple method to rapidly prepare high molecular weight gDNA from Upland cotton, blackgrass, and strawberry suitable for single-molecule sequencing.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.