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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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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 |
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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 |
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