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A method for improving the efficiency of DNA extraction from clotted blood samples

BACKGROUND: The efficient and rapid extraction of high‐quality genomic DNA from clotted blood samples, which normally have a low yield and poor quality, is an important factor in genomic research. The objective of this study was to develop a simple and safe technique for dispersing the blood clots b...

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Autores principales: Mardan‐Nik, Maryam, Saffar Soflaei, Sara, Biabangard‐Zak, Atefeh, Asghari, Mahla, Saljoughian, Sania, Tajbakhsh, Amir, Meshkat, Zahra, Ferns, Gordon A., Pasdar, Alireza, Ghayour‐Mobarhan, Majid
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6642314/
https://www.ncbi.nlm.nih.gov/pubmed/31074532
http://dx.doi.org/10.1002/jcla.22892
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author Mardan‐Nik, Maryam
Saffar Soflaei, Sara
Biabangard‐Zak, Atefeh
Asghari, Mahla
Saljoughian, Sania
Tajbakhsh, Amir
Meshkat, Zahra
Ferns, Gordon A.
Pasdar, Alireza
Ghayour‐Mobarhan, Majid
author_facet Mardan‐Nik, Maryam
Saffar Soflaei, Sara
Biabangard‐Zak, Atefeh
Asghari, Mahla
Saljoughian, Sania
Tajbakhsh, Amir
Meshkat, Zahra
Ferns, Gordon A.
Pasdar, Alireza
Ghayour‐Mobarhan, Majid
author_sort Mardan‐Nik, Maryam
collection PubMed
description BACKGROUND: The efficient and rapid extraction of high‐quality genomic DNA from clotted blood samples, which normally have a low yield and poor quality, is an important factor in genomic research. The objective of this study was to develop a simple and safe technique for dispersing the blood clots by the ball bearing metal shots. Normally, such clot samples may not have an acceptable yield by conventional DNA extraction methods. Also, in the present study, we have further investigated to improve salting‐out DNA extraction methods. METHODS: Initially, 500 µL phosphate‐buffered saline (PBS) (1×) and two ball bearing metal shots were added to each tube of the clotted blood sample and then were gently rotated in an electric laboratory rotator for 1 hour at room temperature (18‐25°C). Genomic DNA was then extracted from samples using a modified salting‐out method and a modified QIAamp(®) DNA Blood Midi Kit and was compared with QIAamp(®) DNA Blood Midi Kit as a control. An assessment of the concentration and quality of the extracted DNA was performed using the UV‐visible spectrophotometer. The isolated DNA proved amenable to PCR amplification and gel electrophoresis. RESULTS: The yield and purity of DNA obtained by these three methods were significantly different (P < 0.001), with a higher yield in the modified salting‐out method. CONCLUSIONS: Our proposed modified salting‐out method is simple and efficient for the isolation of DNA from old blood clot samples. It is both easy to use and is of low cost in routine laboratory tasks.
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spelling pubmed-66423142019-11-12 A method for improving the efficiency of DNA extraction from clotted blood samples Mardan‐Nik, Maryam Saffar Soflaei, Sara Biabangard‐Zak, Atefeh Asghari, Mahla Saljoughian, Sania Tajbakhsh, Amir Meshkat, Zahra Ferns, Gordon A. Pasdar, Alireza Ghayour‐Mobarhan, Majid J Clin Lab Anal Research Articles BACKGROUND: The efficient and rapid extraction of high‐quality genomic DNA from clotted blood samples, which normally have a low yield and poor quality, is an important factor in genomic research. The objective of this study was to develop a simple and safe technique for dispersing the blood clots by the ball bearing metal shots. Normally, such clot samples may not have an acceptable yield by conventional DNA extraction methods. Also, in the present study, we have further investigated to improve salting‐out DNA extraction methods. METHODS: Initially, 500 µL phosphate‐buffered saline (PBS) (1×) and two ball bearing metal shots were added to each tube of the clotted blood sample and then were gently rotated in an electric laboratory rotator for 1 hour at room temperature (18‐25°C). Genomic DNA was then extracted from samples using a modified salting‐out method and a modified QIAamp(®) DNA Blood Midi Kit and was compared with QIAamp(®) DNA Blood Midi Kit as a control. An assessment of the concentration and quality of the extracted DNA was performed using the UV‐visible spectrophotometer. The isolated DNA proved amenable to PCR amplification and gel electrophoresis. RESULTS: The yield and purity of DNA obtained by these three methods were significantly different (P < 0.001), with a higher yield in the modified salting‐out method. CONCLUSIONS: Our proposed modified salting‐out method is simple and efficient for the isolation of DNA from old blood clot samples. It is both easy to use and is of low cost in routine laboratory tasks. John Wiley and Sons Inc. 2019-05-10 /pmc/articles/PMC6642314/ /pubmed/31074532 http://dx.doi.org/10.1002/jcla.22892 Text en © 2019 The Authors. Journal of Clinical Laboratory Analysis Published by Wiley Periodicals, Inc. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Mardan‐Nik, Maryam
Saffar Soflaei, Sara
Biabangard‐Zak, Atefeh
Asghari, Mahla
Saljoughian, Sania
Tajbakhsh, Amir
Meshkat, Zahra
Ferns, Gordon A.
Pasdar, Alireza
Ghayour‐Mobarhan, Majid
A method for improving the efficiency of DNA extraction from clotted blood samples
title A method for improving the efficiency of DNA extraction from clotted blood samples
title_full A method for improving the efficiency of DNA extraction from clotted blood samples
title_fullStr A method for improving the efficiency of DNA extraction from clotted blood samples
title_full_unstemmed A method for improving the efficiency of DNA extraction from clotted blood samples
title_short A method for improving the efficiency of DNA extraction from clotted blood samples
title_sort method for improving the efficiency of dna extraction from clotted blood samples
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6642314/
https://www.ncbi.nlm.nih.gov/pubmed/31074532
http://dx.doi.org/10.1002/jcla.22892
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