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A high throughput DNA extraction method with high yield and quality
BACKGROUND: Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome), and next-generation sequencing directly from sheared...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3441248/ https://www.ncbi.nlm.nih.gov/pubmed/22839646 http://dx.doi.org/10.1186/1746-4811-8-26 |
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author | Xin, Zhanguo Chen, Junping |
author_facet | Xin, Zhanguo Chen, Junping |
author_sort | Xin, Zhanguo |
collection | PubMed |
description | BACKGROUND: Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome), and next-generation sequencing directly from sheared genomic DNA. A variety of DNA preparation methods and commercial kits are available. However, they are either low throughput, low yield, or costly. Here, we describe a method for high throughput genomic DNA isolation from sorghum [Sorghum bicolor (L.) Moench] leaves and dry seeds with high yield, high quality, and affordable cost. RESULTS: We developed a high throughput DNA isolation method by combining a high yield CTAB extraction method with an improved cleanup procedure based on MagAttract kit. The method yielded large quantity and high quality DNA from both lyophilized sorghum leaves and dry seeds. The DNA yield was improved by nearly 30 fold with 4 times less consumption of MagAttract beads. The method can also be used in other plant species, including cotton leaves and pine needles. CONCLUSION: A high throughput system for DNA extraction from sorghum leaves and seeds was developed and validated. The main advantages of the method are low cost, high yield, high quality, and high throughput. One person can process two 96-well plates in a working day at a cost of $0.10 per sample of magnetic beads plus other consumables that other methods will also need. |
format | Online Article Text |
id | pubmed-3441248 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-34412482012-09-14 A high throughput DNA extraction method with high yield and quality Xin, Zhanguo Chen, Junping Plant Methods Methodology BACKGROUND: Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome), and next-generation sequencing directly from sheared genomic DNA. A variety of DNA preparation methods and commercial kits are available. However, they are either low throughput, low yield, or costly. Here, we describe a method for high throughput genomic DNA isolation from sorghum [Sorghum bicolor (L.) Moench] leaves and dry seeds with high yield, high quality, and affordable cost. RESULTS: We developed a high throughput DNA isolation method by combining a high yield CTAB extraction method with an improved cleanup procedure based on MagAttract kit. The method yielded large quantity and high quality DNA from both lyophilized sorghum leaves and dry seeds. The DNA yield was improved by nearly 30 fold with 4 times less consumption of MagAttract beads. The method can also be used in other plant species, including cotton leaves and pine needles. CONCLUSION: A high throughput system for DNA extraction from sorghum leaves and seeds was developed and validated. The main advantages of the method are low cost, high yield, high quality, and high throughput. One person can process two 96-well plates in a working day at a cost of $0.10 per sample of magnetic beads plus other consumables that other methods will also need. BioMed Central 2012-07-28 /pmc/articles/PMC3441248/ /pubmed/22839646 http://dx.doi.org/10.1186/1746-4811-8-26 Text en Copyright ©2012 Xin and Chen; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Xin, Zhanguo Chen, Junping A high throughput DNA extraction method with high yield and quality |
title | A high throughput DNA extraction method with high yield and quality |
title_full | A high throughput DNA extraction method with high yield and quality |
title_fullStr | A high throughput DNA extraction method with high yield and quality |
title_full_unstemmed | A high throughput DNA extraction method with high yield and quality |
title_short | A high throughput DNA extraction method with high yield and quality |
title_sort | high throughput dna extraction method with high yield and quality |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3441248/ https://www.ncbi.nlm.nih.gov/pubmed/22839646 http://dx.doi.org/10.1186/1746-4811-8-26 |
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