A novel approach for extracting DNA from formalin-fixed paraffin-embedded tissue using microwave

Formalin-fixed paraffin-embedded (FFPE) tissue is the gold standard procedure for tissue preservation. However, the extraction of DNA is a cumbersome procedure as the extracted DNA is fragmented because of the cross-linking effect of formalin. Hence, the aim of the present study is to extract DNA fr...

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Veröffentlicht in:Medical journal. Armed Forces India 2020-07, Vol.76 (3), p.307-311
Hauptverfasser: Singh, Hanspal, Narayan, Bhaskar, Urs, Aadithya B., Kumar Polipalli, Sunil, Kumar, Somesh
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container_end_page 311
container_issue 3
container_start_page 307
container_title Medical journal. Armed Forces India
container_volume 76
creator Singh, Hanspal
Narayan, Bhaskar
Urs, Aadithya B.
Kumar Polipalli, Sunil
Kumar, Somesh
description Formalin-fixed paraffin-embedded (FFPE) tissue is the gold standard procedure for tissue preservation. However, the extraction of DNA is a cumbersome procedure as the extracted DNA is fragmented because of the cross-linking effect of formalin. Hence, the aim of the present study is to extract DNA from FFPE tissues using different techniques with a specific objective of comparing the extracted DNA, both quantitatively and qualitatively. Ten samples of FFPE tissues were retrieved from the archives of the Department of Oral Pathology. Total genomic DNA was extracted by different methods which included QIAamp DNA FFPE Tissue Kit, Norgen DNA FFPE Tissue Kit, phenol–chloroform method, mineral oil–based extraction, M/10 NaOH solution method, and microwave method. A 280–base pair sequence was selected for evaluation of downstream amplification. The statistical analysis was performed using unpaired student's t-test to compare the DNA yields and quality obtained by microwave methods with other methods using SPSS software. Total genomic DNA retrieved by the microwave method was superior to other methods both qualitatively and quantitatively. DNA extraction from FFPE tissues is an onerous task as irreversible bonds form between the nucleic acid during fixation which are difficult to break during DNA retrieval. Hence, the microwave method provides good total genomic DNA which gives better downstream results when compared with other methods.
doi_str_mv 10.1016/j.mjafi.2019.02.007
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subjects DNA extraction
Formalin-fixed paraffin-embedded
Microwave
Original
Polymerase Chain Reaction (PCR)
Tissue preservation
title A novel approach for extracting DNA from formalin-fixed paraffin-embedded tissue using microwave
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