Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge
DNA extraction from wastewater sludge (COD 50000 and BOD 25000 mg/l) was conducted using nine different methods normally used for environmental samples including a procedure used in this study and the results obtained were compared. The quality of the differently extracted DNAs was subsequently asse...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Academic Journals
2010
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| Online Access: | http://psasir.upm.edu.my/id/eprint/13672/1/13672.pdf |
| _version_ | 1796968487290142720 |
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| author | Tabatabaei, Meisam Zakaria @ Mamat, Mohd Rafein Abdul Rahim, Raha Abdullah, Norhani Wright, Aandré-Denis G. Shirai, Yoshihito Shamsara, Mehdi Sakai, Kenji Hassan, Mohd Ali |
| author_facet | Tabatabaei, Meisam Zakaria @ Mamat, Mohd Rafein Abdul Rahim, Raha Abdullah, Norhani Wright, Aandré-Denis G. Shirai, Yoshihito Shamsara, Mehdi Sakai, Kenji Hassan, Mohd Ali |
| author_sort | Tabatabaei, Meisam |
| collection | UPM |
| description | DNA extraction from wastewater sludge (COD 50000 and BOD 25000 mg/l) was conducted using nine different methods normally used for environmental samples including a procedure used in this study and the results obtained were compared. The quality of the differently extracted DNAs was subsequently assessed by measuring humic acid concentration, cell lysis efficiency, polymerase chain reaction (PCR) amplification of methanogenic and eubacterial 16S rDNA. The protocol developed in this study was further evaluated by extracting DNA from various high-strength wastewater sludge samples, denaturing gradient gel electrophoresis (DGGE) and fluorescent in situ hybridization (FISH) analyses. The results revealed that great differences existed among the nine procedures and only a few produced satisfactory results when applied to high-strength wastewater sludge. Thermal shock alone was shown inefficient to disrupt the methanogenic cell wall to release the DNA. The method presented in this study (Procedure 9) is generally recommended because of the low concentration of contaminants and its high efficiency despite its simplicity. |
| first_indexed | 2024-03-06T07:29:03Z |
| format | Article |
| id | upm.eprints-13672 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T07:29:03Z |
| publishDate | 2010 |
| publisher | Academic Journals |
| record_format | dspace |
| spelling | upm.eprints-136722017-11-13T03:48:34Z http://psasir.upm.edu.my/id/eprint/13672/ Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge Tabatabaei, Meisam Zakaria @ Mamat, Mohd Rafein Abdul Rahim, Raha Abdullah, Norhani Wright, Aandré-Denis G. Shirai, Yoshihito Shamsara, Mehdi Sakai, Kenji Hassan, Mohd Ali DNA extraction from wastewater sludge (COD 50000 and BOD 25000 mg/l) was conducted using nine different methods normally used for environmental samples including a procedure used in this study and the results obtained were compared. The quality of the differently extracted DNAs was subsequently assessed by measuring humic acid concentration, cell lysis efficiency, polymerase chain reaction (PCR) amplification of methanogenic and eubacterial 16S rDNA. The protocol developed in this study was further evaluated by extracting DNA from various high-strength wastewater sludge samples, denaturing gradient gel electrophoresis (DGGE) and fluorescent in situ hybridization (FISH) analyses. The results revealed that great differences existed among the nine procedures and only a few produced satisfactory results when applied to high-strength wastewater sludge. Thermal shock alone was shown inefficient to disrupt the methanogenic cell wall to release the DNA. The method presented in this study (Procedure 9) is generally recommended because of the low concentration of contaminants and its high efficiency despite its simplicity. Academic Journals 2010-08-02 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/13672/1/13672.pdf Tabatabaei, Meisam and Zakaria @ Mamat, Mohd Rafein and Abdul Rahim, Raha and Abdullah, Norhani and Wright, Aandré-Denis G. and Shirai, Yoshihito and Shamsara, Mehdi and Sakai, Kenji and Hassan, Mohd Ali (2010) Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge. African Journal of Biotechnology, 9 (31). art. no. 61E19D524217. pp. 4926-4937. ISSN 1684–5315 http://www.academicjournals.org/journal/AJB/article-abstract/61E19D524217 10.5897/AJB10.281 |
| spellingShingle | Tabatabaei, Meisam Zakaria @ Mamat, Mohd Rafein Abdul Rahim, Raha Abdullah, Norhani Wright, Aandré-Denis G. Shirai, Yoshihito Shamsara, Mehdi Sakai, Kenji Hassan, Mohd Ali Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge |
| title | Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge |
| title_full | Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge |
| title_fullStr | Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge |
| title_full_unstemmed | Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge |
| title_short | Comparative study of methods for extraction and purification of environmental DNA from high-strength wastewater sludge |
| title_sort | comparative study of methods for extraction and purification of environmental dna from high strength wastewater sludge |
| url | http://psasir.upm.edu.my/id/eprint/13672/1/13672.pdf |
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