Protocol: a simple method for extracting next-generation sequencing quality genomic DNA from recalcitrant plant species

Next-generation sequencing technologies rely on high quality DNA that is suitable for library preparation followed by sequencing. Some plant species store large amounts of phenolics and polysaccharides within their leaf tissue making genomic DNA extraction difficult. While many DNA extraction method...

Full description

Saved in:
Bibliographic Details
Published in:Plant methods 2014-06, Vol.10 (1), p.21-21
Main Authors: Healey, Adam, Furtado, Agnelo, Cooper, Tal, Henry, Robert J
Format: Article
Language:English
Subjects:
Analysis
Coffea
Corymbia
DNA
DNA sequencing
Eucalyptus
genome
high-throughput nucleotide sequencing
leaves
Methodology
Methods
molecular weight
Nucleotide sequencing
phenolic compounds
Polysaccharides
quality control
Quality management
recalcitrant species
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Next-generation sequencing technologies rely on high quality DNA that is suitable for library preparation followed by sequencing. Some plant species store large amounts of phenolics and polysaccharides within their leaf tissue making genomic DNA extraction difficult. While many DNA extraction methods exist that contend with the presence of phenolics and polysaccharides, these methods rely on long incubations, multiple precipitations or commercially available kits to produce high molecular weight and contaminant-free DNA. In this protocol, we describe simple modifications to the established CTAB- based extraction method that allows for reliable isolation of high molecular weight genomic DNA from difficult to isolate plant species Corymbia (a eucalypt) and Coffea (coffee). The simplified protocol does not require multiple clean up steps or commercial based kits, and the isolated DNA passed stringent quality control standards for whole genome sequencing on Illumina HiSeq and TruSeq sequencing platforms.
ISSN:1746-4811
1746-4811
DOI:10.1186/1746-4811-10-21