Cargando…
Improved Method for DNA Extraction and Purification from Tetrahymena pyriformis
Tetrahymena pyriformis (protozoa) is intensely investigated as a model organism, offering numerous advantages in comprehensive and multidisciplinary studies using morphologic or molecular methods. Since DNA extraction is a vital step of any molecular experiment, here a new mixed surfactant (Sodium d...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6632170/ https://www.ncbi.nlm.nih.gov/pubmed/31164619 http://dx.doi.org/10.3390/mps2020040 |
_version_ | 1783435683410477056 |
---|---|
author | El Maaiden, Ezzouhra El Kharrassi, Youssef Essamadi, Abdel Khalid Moustaid, Khadija Nasser, Boubker |
author_facet | El Maaiden, Ezzouhra El Kharrassi, Youssef Essamadi, Abdel Khalid Moustaid, Khadija Nasser, Boubker |
author_sort | El Maaiden, Ezzouhra |
collection | PubMed |
description | Tetrahymena pyriformis (protozoa) is intensely investigated as a model organism, offering numerous advantages in comprehensive and multidisciplinary studies using morphologic or molecular methods. Since DNA extraction is a vital step of any molecular experiment, here a new mixed surfactant (Sodium dodecyl sulfate (SDS) 20%/Triton X-100) was adopted for effective DNA extraction from Tetrahymena pyriformis under an easy, fast protocol. The efficiency of this technique was then compared with three widely-used alternative techniques, namely the Chelex 100 matrix, Ammonium pyrrolidine dithiocarbamate (APD) complex and SDS–chloroform methods. DNA extraction was analyzed by pulsed-field gel electrophoresis, spectral measurement, fluorometry (Qubit), restriction enzyme digestion, and polymerase chain reaction. Data analysis revealed that the quantity and quality of the recovered DNA varied depending on the applied DNA extraction method. The new method (SDS 20%/Triton X-100) was the most efficient for extracting DNA from Tetrahymena pyriformis with high integrity and purity, affordable cost, less time, and suitability for molecular applications. |
format | Online Article Text |
id | pubmed-6632170 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-66321702019-08-19 Improved Method for DNA Extraction and Purification from Tetrahymena pyriformis El Maaiden, Ezzouhra El Kharrassi, Youssef Essamadi, Abdel Khalid Moustaid, Khadija Nasser, Boubker Methods Protoc Protocol Tetrahymena pyriformis (protozoa) is intensely investigated as a model organism, offering numerous advantages in comprehensive and multidisciplinary studies using morphologic or molecular methods. Since DNA extraction is a vital step of any molecular experiment, here a new mixed surfactant (Sodium dodecyl sulfate (SDS) 20%/Triton X-100) was adopted for effective DNA extraction from Tetrahymena pyriformis under an easy, fast protocol. The efficiency of this technique was then compared with three widely-used alternative techniques, namely the Chelex 100 matrix, Ammonium pyrrolidine dithiocarbamate (APD) complex and SDS–chloroform methods. DNA extraction was analyzed by pulsed-field gel electrophoresis, spectral measurement, fluorometry (Qubit), restriction enzyme digestion, and polymerase chain reaction. Data analysis revealed that the quantity and quality of the recovered DNA varied depending on the applied DNA extraction method. The new method (SDS 20%/Triton X-100) was the most efficient for extracting DNA from Tetrahymena pyriformis with high integrity and purity, affordable cost, less time, and suitability for molecular applications. MDPI 2019-05-15 /pmc/articles/PMC6632170/ /pubmed/31164619 http://dx.doi.org/10.3390/mps2020040 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol El Maaiden, Ezzouhra El Kharrassi, Youssef Essamadi, Abdel Khalid Moustaid, Khadija Nasser, Boubker Improved Method for DNA Extraction and Purification from Tetrahymena pyriformis |
title | Improved Method for DNA Extraction and Purification from Tetrahymena pyriformis |
title_full | Improved Method for DNA Extraction and Purification from Tetrahymena pyriformis |
title_fullStr | Improved Method for DNA Extraction and Purification from Tetrahymena pyriformis |
title_full_unstemmed | Improved Method for DNA Extraction and Purification from Tetrahymena pyriformis |
title_short | Improved Method for DNA Extraction and Purification from Tetrahymena pyriformis |
title_sort | improved method for dna extraction and purification from tetrahymena pyriformis |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6632170/ https://www.ncbi.nlm.nih.gov/pubmed/31164619 http://dx.doi.org/10.3390/mps2020040 |
work_keys_str_mv | AT elmaaidenezzouhra improvedmethodfordnaextractionandpurificationfromtetrahymenapyriformis AT elkharrassiyoussef improvedmethodfordnaextractionandpurificationfromtetrahymenapyriformis AT essamadiabdelkhalid improvedmethodfordnaextractionandpurificationfromtetrahymenapyriformis AT moustaidkhadija improvedmethodfordnaextractionandpurificationfromtetrahymenapyriformis AT nasserboubker improvedmethodfordnaextractionandpurificationfromtetrahymenapyriformis |