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Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue

Direct extraction of DNA from the environment has become major importance for molecular analyses for the study of microbial communities in soil and other decomposing agrowaste. The presence of humic substances in Sago pith residue not only results in low DNA quality but also can lead to PCR amplifi...

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Main Authors: Azham, Zulkharnain, Norhafizah, Sidek, Awang Ahmad Sallehin, Awang Husaini
Format: Article
Language:English
Published: Hibiscus Publisher 2014
Subjects:
QD Chemistry
Online Access:http://ir.unimas.my/id/eprint/9509/1/Azham.pdf
http://ir.unimas.my/id/eprint/9509/
http://journal.hibiscuspublisher.com/index.php/JOBIMB/article/view/150
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spelling my.unimas.ir.95092022-01-25T08:38:09Z http://ir.unimas.my/id/eprint/9509/ Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue Azham, Zulkharnain Norhafizah, Sidek Awang Ahmad Sallehin, Awang Husaini QD Chemistry Direct extraction of DNA from the environment has become major importance for molecular analyses for the study of microbial communities in soil and other decomposing agrowaste. The presence of humic substances in Sago pith residue not only results in low DNA quality but also can lead to PCR amplification inhibition which may hinder most molecular studies. Many of the published protocols have been found to be unsuitable to obtained high amount of yield and low of humic acids contamination. This study presents the evaluation of three different methods for extracting total DNA from Sago pith residue. The methods evaluated were enzymatic lysis, glass bead homogenization and freeze-thaw treatment. Each method were evaluated by 260/280 nm absorbance ratio for protein contamination, 260/230 nm absorbance ratio for other contaminants and PCR amplification for molecular work suitability. Among the three methods, freeze-thaw treatment provided the highest yield of DNA, 5.06±0.01 μg/g of Sago pith residue. Nevertheless, all three methods resulted in poor DNA quality which could be used for PCR amplification. Additional steps of agarose electrophoresis and silica column purification were found to be effective for increasing the equality of the extracted DNA and were validated by positive PCR amplifications Hibiscus Publisher 2014 Article NonPeerReviewed text en http://ir.unimas.my/id/eprint/9509/1/Azham.pdf Azham, Zulkharnain and Norhafizah, Sidek and Awang Ahmad Sallehin, Awang Husaini (2014) Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue. Journal of Biochemistry, Microbiology and Biotechnology, 2 (2). pp. 28-30. ISSN 2289-5779 http://journal.hibiscuspublisher.com/index.php/JOBIMB/article/view/150
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic QD Chemistry
spellingShingle QD Chemistry
Azham, Zulkharnain
Norhafizah, Sidek
Awang Ahmad Sallehin, Awang Husaini
Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue
description Direct extraction of DNA from the environment has become major importance for molecular analyses for the study of microbial communities in soil and other decomposing agrowaste. The presence of humic substances in Sago pith residue not only results in low DNA quality but also can lead to PCR amplification inhibition which may hinder most molecular studies. Many of the published protocols have been found to be unsuitable to obtained high amount of yield and low of humic acids contamination. This study presents the evaluation of three different methods for extracting total DNA from Sago pith residue. The methods evaluated were enzymatic lysis, glass bead homogenization and freeze-thaw treatment. Each method were evaluated by 260/280 nm absorbance ratio for protein contamination, 260/230 nm absorbance ratio for other contaminants and PCR amplification for molecular work suitability. Among the three methods, freeze-thaw treatment provided the highest yield of DNA, 5.06±0.01 μg/g of Sago pith residue. Nevertheless, all three methods resulted in poor DNA quality which could be used for PCR amplification. Additional steps of agarose electrophoresis and silica column purification were found to be effective for increasing the equality of the extracted DNA and were validated by positive PCR amplifications
format Article
author Azham, Zulkharnain
Norhafizah, Sidek
Awang Ahmad Sallehin, Awang Husaini
author_facet Azham, Zulkharnain
Norhafizah, Sidek
Awang Ahmad Sallehin, Awang Husaini
author_sort Azham, Zulkharnain
title Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue
title_short Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue
title_full Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue
title_fullStr Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue
title_full_unstemmed Evaluation of Different Methods for Total DNA Extraction from Sago Pith Residue
title_sort evaluation of different methods for total dna extraction from sago pith residue
publisher Hibiscus Publisher
publishDate 2014
url http://ir.unimas.my/id/eprint/9509/1/Azham.pdf
http://ir.unimas.my/id/eprint/9509/
http://journal.hibiscuspublisher.com/index.php/JOBIMB/article/view/150
_version_ 1724078468046520320
score 13.211869

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