Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit

DNA extraction was carried out on 32 medicinal plant samples available in Malaysia using the TriOmicTM extraction kit. Amounts of 0.1 g flowers or young leaves were ground with liquid nitrogen, lysed at 65°C in RY1plus buffer and followed by RNAse treatment. Then, RY2 buffer was added to the samples...

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Main Authors: Ab Rahim, Mohd Hairul, Abu Bakar, Sade, Yiap, Beow Chin, Abdul Rahim, Raha
Format: Article
Language:English
Published: Fundacao de Pesquisas Cientificas de Ribeirao Preto 2011
Online Access:http://psasir.upm.edu.my/id/eprint/22352/1/Genomic%20DNA%20extraction%20from%20medicinal%20plants%20available%20in%20Malaysia%20using%20a%20TriOmicTM%20improved%20extraction%20kit.pdf
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author Ab Rahim, Mohd Hairul
Abu Bakar, Sade
Yiap, Beow Chin
Abdul Rahim, Raha
author_facet Ab Rahim, Mohd Hairul
Abu Bakar, Sade
Yiap, Beow Chin
Abdul Rahim, Raha
author_sort Ab Rahim, Mohd Hairul
collection UPM
description DNA extraction was carried out on 32 medicinal plant samples available in Malaysia using the TriOmicTM extraction kit. Amounts of 0.1 g flowers or young leaves were ground with liquid nitrogen, lysed at 65°C in RY1plus buffer and followed by RNAse treatment. Then, RY2 buffer was added to the samples and mixed completely by vortexing before removal of cell debris by centrifugation. Supernatants were transferred to fresh microcentrifuge tubes and 0.1 volume RY3 buffer was added to each of the transferred supernatant. The mixtures were applied to spin columns followed by a centrifugation step to remove buffers and other residues. Washing step was carried out twice by applying 70% ethanol to the spin columns. Genomic DNA of the samples was recovered by applying 50 μL TE buffer to the membrane of each spin column, followed by a centrifugation step at room temperature. A modification of the TriOmicTM extraction procedure was carried out by adding chloroform:isoamyl alcohol (24:1) steps in the extraction procedure. The genomic DNA extracted from most of the 32 samples showed an increase of total yield when chloroform:isoamyl alcohol (24:1) steps were applied in the TriOmicTM extraction procedure. This preliminary study is very important for molecular studies of medicinal plants available in Malaysia since the DNA extraction can be completed in a shorter period of time (within 1 h) compared to manual extraction, which entails applying phenol, chloroform and ethanol precipitation, and requires 1-2 days to complete.
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spelling upm.eprints-223522015-10-08T08:36:09Z http://psasir.upm.edu.my/id/eprint/22352/ Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit Ab Rahim, Mohd Hairul Abu Bakar, Sade Yiap, Beow Chin Abdul Rahim, Raha DNA extraction was carried out on 32 medicinal plant samples available in Malaysia using the TriOmicTM extraction kit. Amounts of 0.1 g flowers or young leaves were ground with liquid nitrogen, lysed at 65°C in RY1plus buffer and followed by RNAse treatment. Then, RY2 buffer was added to the samples and mixed completely by vortexing before removal of cell debris by centrifugation. Supernatants were transferred to fresh microcentrifuge tubes and 0.1 volume RY3 buffer was added to each of the transferred supernatant. The mixtures were applied to spin columns followed by a centrifugation step to remove buffers and other residues. Washing step was carried out twice by applying 70% ethanol to the spin columns. Genomic DNA of the samples was recovered by applying 50 μL TE buffer to the membrane of each spin column, followed by a centrifugation step at room temperature. A modification of the TriOmicTM extraction procedure was carried out by adding chloroform:isoamyl alcohol (24:1) steps in the extraction procedure. The genomic DNA extracted from most of the 32 samples showed an increase of total yield when chloroform:isoamyl alcohol (24:1) steps were applied in the TriOmicTM extraction procedure. This preliminary study is very important for molecular studies of medicinal plants available in Malaysia since the DNA extraction can be completed in a shorter period of time (within 1 h) compared to manual extraction, which entails applying phenol, chloroform and ethanol precipitation, and requires 1-2 days to complete. Fundacao de Pesquisas Cientificas de Ribeirao Preto 2011 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/22352/1/Genomic%20DNA%20extraction%20from%20medicinal%20plants%20available%20in%20Malaysia%20using%20a%20TriOmicTM%20improved%20extraction%20kit.pdf Ab Rahim, Mohd Hairul and Abu Bakar, Sade and Yiap, Beow Chin and Abdul Rahim, Raha (2011) Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit. Genetics and Molecular Research, 10 (4). pp. 2757-2764. ISSN 1676-5680 http://www.geneticsmr.com/articles/1367 10.4238/2011.November.8.1
spellingShingle Ab Rahim, Mohd Hairul
Abu Bakar, Sade
Yiap, Beow Chin
Abdul Rahim, Raha
Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit
title Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit
title_full Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit
title_fullStr Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit
title_full_unstemmed Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit
title_short Genomic DNA extraction from medicinal plants available in Malaysia using a TriOmicTM improved extraction kit
title_sort genomic dna extraction from medicinal plants available in malaysia using a triomictm improved extraction kit
url http://psasir.upm.edu.my/id/eprint/22352/1/Genomic%20DNA%20extraction%20from%20medicinal%20plants%20available%20in%20Malaysia%20using%20a%20TriOmicTM%20improved%20extraction%20kit.pdf
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AT yiapbeowchin genomicdnaextractionfrommedicinalplantsavailableinmalaysiausingatriomictmimprovedextractionkit
AT abdulrahimraha genomicdnaextractionfrommedicinalplantsavailableinmalaysiausingatriomictmimprovedextractionkit