Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: El-Ashram, Saeed, Al Nasr, Ibrahim, Suo, Xun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2016
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
_version_ 1782516694253568000
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