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Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method

In case of studies associated with human genetics, genomics, and pharmacogenetics the genomic DNA is extracted from the buccal cells, whole blood etc. Several methods are exploited by the researchers to extract DNA from the whole blood. One of these methods, which utilizes cell lysis and proteolytic...

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Detalles Bibliográficos
Autores principales: Qamar, Wajhul, Khan, Mohammad Rashid, Arafah, Azher
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6169501/
https://www.ncbi.nlm.nih.gov/pubmed/30294214
http://dx.doi.org/10.1016/j.sjbs.2016.09.016
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author Qamar, Wajhul
Khan, Mohammad Rashid
Arafah, Azher
author_facet Qamar, Wajhul
Khan, Mohammad Rashid
Arafah, Azher
author_sort Qamar, Wajhul
collection PubMed
description In case of studies associated with human genetics, genomics, and pharmacogenetics the genomic DNA is extracted from the buccal cells, whole blood etc. Several methods are exploited by the researchers to extract DNA from the whole blood. One of these methods, which utilizes cell lysis and proteolytic properties of sodium dodecyl sulfate (SDS) and proteinase K respectively, might also be called SDS-PK method. It does not include any hazardous chemicals such as phenol or chloroform and is inexpensive. However, several researchers report the same method with different formulas and conditions. During our experiments with whole blood DNA extraction we experienced problems such as protein contamination, DNA purity and yield when followed some SDS-PK protocols reported elsewhere. A260/A280 and A260/A230 ratios along with PCR amplification give a clear idea about the procedure that was followed to extract the DNA. In an effort to increase the DNA purity from human whole blood, we pointed out some steps of the protocol that play a crucial role in determining the extraction of high quality DNA.
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spelling pubmed-61695012018-10-05 Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method Qamar, Wajhul Khan, Mohammad Rashid Arafah, Azher Saudi J Biol Sci Article In case of studies associated with human genetics, genomics, and pharmacogenetics the genomic DNA is extracted from the buccal cells, whole blood etc. Several methods are exploited by the researchers to extract DNA from the whole blood. One of these methods, which utilizes cell lysis and proteolytic properties of sodium dodecyl sulfate (SDS) and proteinase K respectively, might also be called SDS-PK method. It does not include any hazardous chemicals such as phenol or chloroform and is inexpensive. However, several researchers report the same method with different formulas and conditions. During our experiments with whole blood DNA extraction we experienced problems such as protein contamination, DNA purity and yield when followed some SDS-PK protocols reported elsewhere. A260/A280 and A260/A230 ratios along with PCR amplification give a clear idea about the procedure that was followed to extract the DNA. In an effort to increase the DNA purity from human whole blood, we pointed out some steps of the protocol that play a crucial role in determining the extraction of high quality DNA. Elsevier 2017-11 2016-09-10 /pmc/articles/PMC6169501/ /pubmed/30294214 http://dx.doi.org/10.1016/j.sjbs.2016.09.016 Text en © 2016 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Qamar, Wajhul
Khan, Mohammad Rashid
Arafah, Azher
Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method
title Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method
title_full Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method
title_fullStr Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method
title_full_unstemmed Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method
title_short Optimization of conditions to extract high quality DNA for PCR analysis from whole blood using SDS-proteinase K method
title_sort optimization of conditions to extract high quality dna for pcr analysis from whole blood using sds-proteinase k method
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6169501/
https://www.ncbi.nlm.nih.gov/pubmed/30294214
http://dx.doi.org/10.1016/j.sjbs.2016.09.016
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