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/9509/ http://ir.unimas.my/9509/ http://ir.unimas.my/9509/1/NO%2068%20Evaluation%20of%20Different%20Methods%20for%20Total%20DNA%20Extraction%20from%20Sago%20%28abstract%29.pdf

id	unimas-9509
recordtype	eprints
spelling	unimas-95092017-03-01T01:40:51Z http://ir.unimas.my/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/9509/1/NO%2068%20Evaluation%20of%20Different%20Methods%20for%20Total%20DNA%20Extraction%20from%20Sago%20%28abstract%29.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
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Malaysia Sarawak
building	UNIMAS Institutional Repository
collection	Online Access
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/9509/ http://ir.unimas.my/9509/ http://ir.unimas.my/9509/1/NO%2068%20Evaluation%20of%20Different%20Methods%20for%20Total%20DNA%20Extraction%20from%20Sago%20%28abstract%29.pdf
first_indexed	2018-09-06T15:40:18Z
last_indexed	2018-09-06T15:40:18Z
_version_	1610873221941297152

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

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