Cargando…

Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize

BACKGROUND: The world’s top three cereals, based on their monetary value, are rice, wheat, and corn. In cereal crops, DNA extraction is difficult owing to rigid non-cellulose components in the cell wall of leaves and high starch and protein content in grains. The advanced techniques in molecular bio...

Descripción completa

Detalles Bibliográficos
Autores principales: Abdel-Latif, Amani, Osman, Gamal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5209869/
https://www.ncbi.nlm.nih.gov/pubmed/28053646
http://dx.doi.org/10.1186/s13007-016-0152-4
_version_ 1782490810128793600
author Abdel-Latif, Amani
Osman, Gamal
author_facet Abdel-Latif, Amani
Osman, Gamal
author_sort Abdel-Latif, Amani
collection PubMed
description BACKGROUND: The world’s top three cereals, based on their monetary value, are rice, wheat, and corn. In cereal crops, DNA extraction is difficult owing to rigid non-cellulose components in the cell wall of leaves and high starch and protein content in grains. The advanced techniques in molecular biology require pure and quick extraction of DNA. The majority of existing DNA extraction methods rely on long incubation and multiple precipitations or commercially available kits to produce contaminant-free high molecular weight DNA. RESULTS: In this study, we compared three different methods used for the isolation of high-quality genomic DNA from the grains of cereal crop, Zea mays, with minor modifications. The DNA from the grains of two maize hybrids, M10 and M321, was extracted using extraction methods DNeasy Qiagen Plant Mini Kit, CTAB-method (with/without 1% PVP) and modified Mericon extraction. Genes coding for 45S ribosomal RNA are organized in tandem arrays of up to several thousand copies and contain codes for 18S, 5.8S and 26S rRNA units separated by internal transcribed spacers ITS1 and ITS2. While the rRNA units are evolutionary conserved, ITS regions show high level of interspecific divergence and have been used frequently in genetic diversity and phylogenetic studies. In this study, the genomic DNA was then amplified with PCR using primers specific for ITS gene. PCR products were then visualized on agarose gel. CONCLUSION: The modified Mericon extraction method was found to be the most efficient DNA extraction method, capable to provide high DNA yields with better quality, affordable cost and less time.
format Online
Article
Text
id pubmed-5209869
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-52098692017-01-04 Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize Abdel-Latif, Amani Osman, Gamal Plant Methods Research BACKGROUND: The world’s top three cereals, based on their monetary value, are rice, wheat, and corn. In cereal crops, DNA extraction is difficult owing to rigid non-cellulose components in the cell wall of leaves and high starch and protein content in grains. The advanced techniques in molecular biology require pure and quick extraction of DNA. The majority of existing DNA extraction methods rely on long incubation and multiple precipitations or commercially available kits to produce contaminant-free high molecular weight DNA. RESULTS: In this study, we compared three different methods used for the isolation of high-quality genomic DNA from the grains of cereal crop, Zea mays, with minor modifications. The DNA from the grains of two maize hybrids, M10 and M321, was extracted using extraction methods DNeasy Qiagen Plant Mini Kit, CTAB-method (with/without 1% PVP) and modified Mericon extraction. Genes coding for 45S ribosomal RNA are organized in tandem arrays of up to several thousand copies and contain codes for 18S, 5.8S and 26S rRNA units separated by internal transcribed spacers ITS1 and ITS2. While the rRNA units are evolutionary conserved, ITS regions show high level of interspecific divergence and have been used frequently in genetic diversity and phylogenetic studies. In this study, the genomic DNA was then amplified with PCR using primers specific for ITS gene. PCR products were then visualized on agarose gel. CONCLUSION: The modified Mericon extraction method was found to be the most efficient DNA extraction method, capable to provide high DNA yields with better quality, affordable cost and less time. BioMed Central 2017-01-03 /pmc/articles/PMC5209869/ /pubmed/28053646 http://dx.doi.org/10.1186/s13007-016-0152-4 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Abdel-Latif, Amani
Osman, Gamal
Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize
title Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize
title_full Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize
title_fullStr Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize
title_full_unstemmed Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize
title_short Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize
title_sort comparison of three genomic dna extraction methods to obtain high dna quality from maize
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5209869/
https://www.ncbi.nlm.nih.gov/pubmed/28053646
http://dx.doi.org/10.1186/s13007-016-0152-4
work_keys_str_mv AT abdellatifamani comparisonofthreegenomicdnaextractionmethodstoobtainhighdnaqualityfrommaize
AT osmangamal comparisonofthreegenomicdnaextractionmethodstoobtainhighdnaqualityfrommaize