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Nucleic acid protocols: Extraction and optimization
Yield and quality are fundamental features for any researchers during nucleic acid extraction. Here, we describe a simplified, semi-unified, effective, and toxic material free protocol for extracting DNA and RNA from different prokaryotic and eukaryotic sources exploiting the physical and chemical p...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5361071/ https://www.ncbi.nlm.nih.gov/pubmed/28352552 http://dx.doi.org/10.1016/j.btre.2016.10.001 |
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author | El-Ashram, Saeed Al Nasr, Ibrahim Suo, Xun |
author_facet | El-Ashram, Saeed Al Nasr, Ibrahim Suo, Xun |
author_sort | El-Ashram, Saeed |
collection | PubMed |
description | Yield and quality are fundamental features for any researchers during nucleic acid extraction. Here, we describe a simplified, semi-unified, effective, and toxic material free protocol for extracting DNA and RNA from different prokaryotic and eukaryotic sources exploiting the physical and chemical properties of nucleic acids. Furthermore, this protocol showed that DNA and RNA are under triple protection (i.e. EDTA, SDS and NaCl) during lysis step, and this environment is improper for RNase to have DNA liberated of RNA and even for DNase to degrade the DNA. Therefore, the complete removal of RNA under RNase influence is achieved when RNase is added after DNA extraction, which gives optimal quality with any protocols. Similarly, DNA contamination in an isolated RNA is degraded by DNase to obtain high-quality RNA. Our protocol is the protocol of choice in terms of simplicity, recovery time, environmental safety, amount, purity, PCR and RT-PCR applicability. |
format | Online Article Text |
id | pubmed-5361071 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-53610712017-03-28 Nucleic acid protocols: Extraction and optimization El-Ashram, Saeed Al Nasr, Ibrahim Suo, Xun Biotechnol Rep (Amst) Article Yield and quality are fundamental features for any researchers during nucleic acid extraction. Here, we describe a simplified, semi-unified, effective, and toxic material free protocol for extracting DNA and RNA from different prokaryotic and eukaryotic sources exploiting the physical and chemical properties of nucleic acids. Furthermore, this protocol showed that DNA and RNA are under triple protection (i.e. EDTA, SDS and NaCl) during lysis step, and this environment is improper for RNase to have DNA liberated of RNA and even for DNase to degrade the DNA. Therefore, the complete removal of RNA under RNase influence is achieved when RNase is added after DNA extraction, which gives optimal quality with any protocols. Similarly, DNA contamination in an isolated RNA is degraded by DNase to obtain high-quality RNA. Our protocol is the protocol of choice in terms of simplicity, recovery time, environmental safety, amount, purity, PCR and RT-PCR applicability. Elsevier 2016-10-05 /pmc/articles/PMC5361071/ /pubmed/28352552 http://dx.doi.org/10.1016/j.btre.2016.10.001 Text en © 2016 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article El-Ashram, Saeed Al Nasr, Ibrahim Suo, Xun Nucleic acid protocols: Extraction and optimization |
title | Nucleic acid protocols: Extraction and optimization |
title_full | Nucleic acid protocols: Extraction and optimization |
title_fullStr | Nucleic acid protocols: Extraction and optimization |
title_full_unstemmed | Nucleic acid protocols: Extraction and optimization |
title_short | Nucleic acid protocols: Extraction and optimization |
title_sort | nucleic acid protocols: extraction and optimization |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5361071/ https://www.ncbi.nlm.nih.gov/pubmed/28352552 http://dx.doi.org/10.1016/j.btre.2016.10.001 |
work_keys_str_mv | AT elashramsaeed nucleicacidprotocolsextractionandoptimization AT alnasribrahim nucleicacidprotocolsextractionandoptimization AT suoxun nucleicacidprotocolsextractionandoptimization |