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@ARTICLE{Vilanova:878413,
author = {Vilanova, Santiago and Alonso, David and Gramazio, Pietro
and Plazas, Mariola and García-Fortea, Edgar and Ferrante,
Paola and Schmidt, Maximilian and Díez, María José and
Usadel, Björn and Giuliano, Giovanni and Prohens, Jaime},
title = {{SILEX}: a fast and inexpensive high-quality {DNA}
extraction method suitable for multiple sequencing platforms
and recalcitrant plant species},
journal = {Plant methods},
volume = {16},
number = {1},
issn = {1746-4811},
address = {London},
publisher = {BioMed Central},
reportid = {FZJ-2020-02843},
pages = {110},
year = {2020},
abstract = {Background: The use of sequencing and genotyping platforms
has undergone dramatic improvements, enablingthe generation
of a wealth of genomic information. Despite this progress,
the availability of high-quality genomicDNA (gDNA) in
sufcient concentrations is often a main limitation,
especially for third-generation sequencing plat‑forms. A
variety of DNA extraction methods and commercial kits are
available. However, many of these are costly andfrequently
give either low yield or low-quality DNA, inappropriate for
next generation sequencing (NGS) platforms.Here, we describe
a fast and inexpensive DNA extraction method (SILEX)
applicable to a wide range of plant speciesand
tissues.Results: SILEX is a high-throughput DNA extraction
protocol, based on the standard CTAB method with a DNA
silicamatrix recovery, which allows obtaining NGS-quality
high molecular weight genomic plant DNA free of
inhibitorycompounds. SILEX was compared with a standard CTAB
extraction protocol and a common commercial extraction kitin
a variety of species, including recalcitrant ones, from
diferent families. In comparison with the other methods,
SILEXyielded DNA in higher concentrations and of higher
quality. Manual extraction of 48 samples can be done in 96
minby one person at a cost of 0.12 €/sample of reagents
and consumables. Hundreds of tomato gDNA samples
obtainedwith either SILEX or the commercial kit were
successfully genotyped with Single Primer Enrichment
Technology(SPET) with the Illumina HiSeq 2500 platform.
Furthermore, DNA extracted from Solanum elaeagnifolium using
thisprotocol was assessed by Pulsed-feld gel electrophoresis
(PFGE), obtaining a suitable size ranges for most
sequencingplatforms that required high-molecular-weight DNA
such as Nanopore or PacBio.Conclusions: A high-throughput,
fast and inexpensive DNA extraction protocol was developed
and validated for awide variety of plants and tissues. SILEX
ofers an easy, scalable, efcient and inexpensive way to
extract DNA for vari‑ous next-generation sequencing
applications including SPET and Nanopore among
others.Keywords: DNA extraction, CTAB protocol, Silica
matrix, Contaminant-free DNA, High-molecular-weight DNA,
Nextgeneration sequencing, High-throughput genotyping,
Recalcitrant species, SPET, Nanopore},
cin = {IBG-4},
ddc = {570},
cid = {I:(DE-Juel1)IBG-4-20200403},
pnm = {582 - Plant Science (POF3-582) / 583 - Innovative
Synergisms (POF3-583)},
pid = {G:(DE-HGF)POF3-582 / G:(DE-HGF)POF3-583},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:32793297},
UT = {WOS:000561439100002},
doi = {10.1186/s13007-020-00652-y},
url = {https://juser.fz-juelich.de/record/878413},
}