ArticleNature protocols2012
Preparation of megabase-sized DNA from a variety of organisms using the nuclei method for advanced genomics research.
Article in Nature protocols, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 82 papers.
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Who cites it
82 citing papers in PubMed.
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- Structure and evolution of the Forsythieae genome elucidated by chromosome-level genome comparison of Abeliophyllum distichum and Forsythia ovata (Oleaceae).Communications biology · 2025Article
- Chromosome-level assemblies of the endemic Korean species Abeliophyllum distichum and Forsythia ovata.Scientific data · 2024Article
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- Gapless assembly of complete human and plant chromosomes using only nanopore sequencing.Genome research · 2024Article
- A CRISPR-Cas9-Mediated Large-Fragment Assembly Method for Cloning Genomes and Biosynthetic Gene Cluster.Microorganisms · 2024Article
- Chromosome-level genome assembly of Korean holoparasitic plants, Orobanche coerulescens.Scientific data · 2024Article
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- Gapless assembly of complete human and plant chromosomes using only nanopore sequencing.bioRxiv : the preprint server for biology · 2024Article
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- Dissection of a rapidly evolving wheat resistance gene cluster by long-read genome sequencing accelerated the cloning of Pm69.Plant communications · 2024Article
22 more citing papers are in PubMed but not listed here.
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6 authors.
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Abstract
Megabase-sized DNA is crucial to modern genomics research of all organisms. Among the preparation methods developed, the nuclei method is the simplest and most widely used for preparing high-quality megabase-sized DNA from divergent organisms. In this method, nuclei are first isolated by physically grinding the source tissues. The nontarget cytoplast organellar genomes and metabolites are removed by centrifugation and washing, thus maximizing the utility of the method and substantially improving the digestibility and clonability of the resultant DNA. The nuclei are then embedded in an agarose matrix containing numerous pores, allowing the access of restriction enzymes while preventing the DNA from physical shearing. DNA is extracted from the nuclei, purified and subsequently manipulated in the agarose matrix. Here we describe the nuclei method that we have successfully used to prepare high-quality megabase-sized DNA from hundreds of plant, animal, fish, insect, algal and microbial species. The entire protocol takes ∼3 d.
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