RustFungiV1, Main, Exploration, bibRecord, 000542

Extraction of High Molecular Weight DNA from Fungal Rust Spores for Long Read Sequencing.

Identifieur interne : 000542 ( Main/Exploration ); précédent : 000541; suivant : 000543

Extraction of High Molecular Weight DNA from Fungal Rust Spores for Long Read Sequencing.

Auteurs : Benjamin Schwessinger [Australie] ; John P. Rathjen [Australie]

Source :

Methods in molecular biology (Clifton, N.J.) [ 1940-6029 ] ; 2017.

RBID : pubmed:28856640

Descripteurs français

KwdFr :
- ADN fongique (composition chimique), ADN fongique (génétique), ADN fongique (isolement et purification), Analyse de séquence d'ADN (méthodes), Basidiomycota (composition chimique), Basidiomycota (génétique), Fractionnement chimique (méthodes), Maladies des plantes (microbiologie), Masse moléculaire (MeSH), Spores fongiques (composition chimique), Spores fongiques (génétique), Triticum (microbiologie).
MESH :
- composition chimique : ADN fongique, Basidiomycota, Spores fongiques.
- génétique : ADN fongique, Basidiomycota, Spores fongiques.
- isolement et purification : ADN fongique.
- microbiologie : Maladies des plantes, Triticum.
- méthodes : Analyse de séquence d'ADN, Fractionnement chimique.
- Masse moléculaire.

English descriptors

KwdEn :
- Basidiomycota (chemistry), Basidiomycota (genetics), Chemical Fractionation (methods), DNA, Fungal (chemistry), DNA, Fungal (genetics), DNA, Fungal (isolation & purification), Molecular Weight (MeSH), Plant Diseases (microbiology), Sequence Analysis, DNA (methods), Spores, Fungal (chemistry), Spores, Fungal (genetics), Triticum (microbiology).
MESH :
- chemical , chemistry : DNA, Fungal.
- chemistry : Basidiomycota, Spores, Fungal.
- genetics : Basidiomycota, DNA, Fungal, Spores, Fungal.
- chemical , isolation & purification : DNA, Fungal.
- methods : Chemical Fractionation, Sequence Analysis, DNA.
- microbiology : Plant Diseases, Triticum.
- Molecular Weight.

Abstract

Wheat rust fungi are complex organisms with a complete life cycle that involves two different host plants and five different spore types. During the asexual infection cycle on wheat, rusts produce massive amounts of dikaryotic urediniospores. These spores are dikaryotic (two nuclei) with each nucleus containing one haploid genome. This dikaryotic state is likely to contribute to their evolutionary success, making them some of the major wheat pathogens globally. Despite this, most published wheat rust genomes are highly fragmented and contain very little haplotype-specific sequence information. Current long-read sequencing technologies hold great promise to provide more contiguous and haplotype-phased genome assemblies. Long reads are able to span repetitive regions and phase structural differences between the haplomes. This increased genome resolution enables the identification of complex loci and the study of genome evolution beyond simple nucleotide polymorphisms. Long-read technologies require pure high molecular weight DNA as an input for sequencing. Here, we describe a DNA extraction protocol for rust spores that yields pure double-stranded DNA molecules with molecular weight of >50 kilo-base pairs (kbp). The isolated DNA is of sufficient purity for PacBio long-read sequencing, but may require additional purification for other sequencing technologies such as Nanopore and 10× Genomics.

DOI: 10.1007/978-1-4939-7249-4_5
PubMed: 28856640

Affiliations:

Australie

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Le document en format XML

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<term>Analyse de séquence d'ADN (méthodes)</term>
<term>Basidiomycota (composition chimique)</term>
<term>Basidiomycota (génétique)</term>
<term>Fractionnement chimique (méthodes)</term>
<term>Maladies des plantes (microbiologie)</term>
<term>Masse moléculaire (MeSH)</term>
<term>Spores fongiques (composition chimique)</term>
<term>Spores fongiques (génétique)</term>
<term>Triticum (microbiologie)</term>
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<front><div type="abstract" xml:lang="en">Wheat rust fungi are complex organisms with a complete life cycle that involves two different host plants and five different spore types. During the asexual infection cycle on wheat, rusts produce massive amounts of dikaryotic urediniospores. These spores are dikaryotic (two nuclei) with each nucleus containing one haploid genome. This dikaryotic state is likely to contribute to their evolutionary success, making them some of the major wheat pathogens globally. Despite this, most published wheat rust genomes are highly fragmented and contain very little haplotype-specific sequence information. Current long-read sequencing technologies hold great promise to provide more contiguous and haplotype-phased genome assemblies. Long reads are able to span repetitive regions and phase structural differences between the haplomes. This increased genome resolution enables the identification of complex loci and the study of genome evolution beyond simple nucleotide polymorphisms. Long-read technologies require pure high molecular weight DNA as an input for sequencing. Here, we describe a DNA extraction protocol for rust spores that yields pure double-stranded DNA molecules with molecular weight of >50 kilo-base pairs (kbp). The isolated DNA is of sufficient purity for PacBio long-read sequencing, but may require additional purification for other sequencing technologies such as Nanopore and 10× Genomics.</div>
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<Keyword MajorTopicYN="Y">Puccinia graminis f. sp. tritici</Keyword>
<Keyword MajorTopicYN="Y">Puccinia striiformis f. sp. tritici</Keyword>
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Extraction of High Molecular Weight DNA from Fungal Rust Spores for Long Read Sequencing.

Extraction of High Molecular Weight DNA from Fungal Rust Spores for Long Read Sequencing.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki