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

Maryam Mardan‐Nik; Sara Saffar Soflaei; Atefeh Biabangard‐Zak; Mahla Asghari; Sania Saljoughian; Amir Tajbakhsh; Zahra Meshkat; Gordon A. Ferns; Alireza Pasdar; Majid Ghayour‐Mobarhan

doi:10.1002/jcla.22892

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

Maryam Mardan‐Nik, Sara Saffar Soflaei, Atefeh Biabangard‐Zak, Mahla Asghari, Sania Saljoughian, Amir Tajbakhsh, Zahra Meshkat, Gordon A. Ferns, Alireza Pasdar^* (Corresponding Author), Majid Ghayour‐Mobarhan^* (Corresponding Author)

^*Corresponding author for this work

Applied Medicine

Research output: Contribution to journal › Article › peer-review

21 Citations (Scopus)

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Abstract

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.

Original language	English
Article number	e22892
Number of pages	7
Journal	Journal of Clinical Laboratory Analysis
Volume	33
Issue number	6
Early online date	10 May 2019
DOIs	https://doi.org/10.1002/jcla.22892
Publication status	Published - Jul 2019

Bibliographical note

Funding information:
This study was supported by a grant from the Research Council of the Mashhad University of Medical Sciences (Grant No: 931680).

The authors would like to thank Dr. Hossein Eshghi at Department of Chemistry, Faculty of Science, the Ferdowsi University of Mashhad for his assistance in the experiment and Mohammad Sadegh Khorami who contributed to this study. We are also particularly grateful to the Research Council of the Mashhad University of Medical Sciences (MUMS) for the financial support of this study

Keywords

clotted blood
DNA extraction
frozen
salting out

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Cite this

@article{661b9f11847b414d9c60cbdd92f8f4ac,

title = "A method for improving the efficiency of DNA extraction from clotted blood samples",

abstract = "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{\textregistered} DNA Blood Midi Kit and was compared with QIAamp{\textregistered} 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. ",

keywords = "clotted blood, DNA extraction, frozen, salting out",

author = "Maryam Mardan‐Nik and Soflaei, {Sara Saffar} and Atefeh Biabangard‐Zak and Mahla Asghari and Sania Saljoughian and Amir Tajbakhsh and Zahra Meshkat and Ferns, {Gordon A.} and Alireza Pasdar and Majid Ghayour‐Mobarhan",

note = "Funding information: This study was supported by a grant from the Research Council of the Mashhad University of Medical Sciences (Grant No: 931680). The authors would like to thank Dr. Hossein Eshghi at Department of Chemistry, Faculty of Science, the Ferdowsi University of Mashhad for his assistance in the experiment and Mohammad Sadegh Khorami who contributed to this study. We are also particularly grateful to the Research Council of the Mashhad University of Medical Sciences (MUMS) for the financial support of this study",

year = "2019",

month = jul,

doi = "10.1002/jcla.22892",

language = "English",

volume = "33",

journal = "Journal of Clinical Laboratory Analysis",

issn = "1098-2825",

publisher = "Wiley-Liss Inc.",

number = "6",

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T1 - A method for improving the efficiency of DNA extraction from clotted blood samples

AU - Mardan‐Nik, Maryam

AU - Soflaei, Sara Saffar

AU - Biabangard‐Zak, Atefeh

AU - Asghari, Mahla

AU - Saljoughian, Sania

AU - Tajbakhsh, Amir

AU - Meshkat, Zahra

AU - Ferns, Gordon A.

AU - Pasdar, Alireza

AU - Ghayour‐Mobarhan, Majid

N1 - Funding information: This study was supported by a grant from the Research Council of the Mashhad University of Medical Sciences (Grant No: 931680). The authors would like to thank Dr. Hossein Eshghi at Department of Chemistry, Faculty of Science, the Ferdowsi University of Mashhad for his assistance in the experiment and Mohammad Sadegh Khorami who contributed to this study. We are also particularly grateful to the Research Council of the Mashhad University of Medical Sciences (MUMS) for the financial support of this study

PY - 2019/7

Y1 - 2019/7

N2 - 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.

AB - 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.

KW - clotted blood

KW - DNA extraction

KW - frozen

KW - salting out

U2 - 10.1002/jcla.22892

DO - 10.1002/jcla.22892

M3 - Article

SN - 1098-2825

VL - 33

JO - Journal of Clinical Laboratory Analysis

JF - Journal of Clinical Laboratory Analysis

IS - 6

M1 - e22892

ER -

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

Abstract

Bibliographical note

Keywords

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