Nigerian Medical Journal

ORIGINAL ARTICLE
Year
: 2016  |  Volume : 57  |  Issue : 4  |  Page : 199--203

The effect of deoxyribonucleic acid extraction methods from lymphoid tissue on the purity, content, and amplifying ability


Hossein Ayatollahi1, Mohammad Hadi Sadeghian1, Mohammad Reza Keramati2, Ali Ayatollahi3, Arezoo Shajiei3, Maryam Sheikhi3, Samane Bakhshi3 
1 Department of Hematopathology and Blood Banking, Cancer Molecular Pathology Research Center, Faculty of Medicine, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran
2 Department of Internal Medicine, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
3 Cancer Molecular Pathology Research Center, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran

Correspondence Address:
Maryam Sheikhi
Cancer Molecular Pathology Research Center, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad
Iran

Background: Nowadays, definitive diagnosis of numerous diseases is based on the genetic and molecular findings. Therefore, preparation of fundamental materials for these evaluations is necessary. Deoxyribonucleic acid (DNA) is the first material for the molecular pathology and genetic analysis, and better results need more pure DNA. Furthermore, higher concentration of achieved DNA causes better results and higher amplifying ability for subsequent steps. We aim to evaluate five DNA extraction methods to compare DNA intimacy including purity, concentration, and amplifying ability with each other. Materials and Methods: The lymphoid tissue DNA was extracted from formalin-fixed, paraffin embedded (FFPE) tissue through five different methods including phenol-chloroform as the reference method, DNA isolation kit (QIAamp DNA FFPE Tissue Kit, Qiagen, Germany), proteinase K and xylol extraction and heat alkaline plus mineral oil extraction as authorship innovative method. Finally, polymerase chain reaction (PCR) and real-time PCR method were assessed to compare each following method consider to DNA purity and its concentration. Results: Among five different applied methods, the highest mean of DNA purity was related to heat alkaline method. Moreover, the highest mean of DNA concentration was related to heat alkaline plus mineral oil. Furthermore, the best result in quantitative PCR was in proteinase K method that had the lowest cycle threshold averages among the other extraction methods. Conclusion: We concluded that our innovative method for DNA extraction (heat alkaline plus mineral oil) achieved high DNA purity and concentration.


How to cite this article:
Ayatollahi H, Sadeghian MH, Keramati MR, Ayatollahi A, Shajiei A, Sheikhi M, Bakhshi S. The effect of deoxyribonucleic acid extraction methods from lymphoid tissue on the purity, content, and amplifying ability.Niger Med J 2016;57:199-203


How to cite this URL:
Ayatollahi H, Sadeghian MH, Keramati MR, Ayatollahi A, Shajiei A, Sheikhi M, Bakhshi S. The effect of deoxyribonucleic acid extraction methods from lymphoid tissue on the purity, content, and amplifying ability. Niger Med J [serial online] 2016 [cited 2022 Jan 19 ];57:199-203
Available from: https://www.nigeriamedj.com/article.asp?issn=0300-1652;year=2016;volume=57;issue=4;spage=199;epage=203;aulast=Ayatollahi;type=0