A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies

A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies

High RNA quality, integrity and reproducibility are essential for functional genomics study involving library preparation and sequencing using next generation sequencing (NGS) technologies. The medicinal herbaceous plant, Impatiens balsamina, possess a rich source of diverse bioactive compounds in...

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Main Authors: Foong, Lian Chee, Siong Hock Ho, Anthony Hock Ho, Lim, Yang Mooi, Tam, Sheh May
Format: Article
Language:English
Published: Penerbit Universiti Kebangsaan Malaysia 2017
Online Access:http://journalarticle.ukm.my/12336/
http://journalarticle.ukm.my/12336/
http://journalarticle.ukm.my/12336/1/46_02_11.pdf
id ukm-12336
recordtype eprints
spelling ukm-123362018-11-16T21:27:28Z http://journalarticle.ukm.my/12336/ A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies Foong, Lian Chee Siong Hock Ho, Anthony Hock Ho Lim, Yang Mooi Tam, Sheh May High RNA quality, integrity and reproducibility are essential for functional genomics study involving library preparation and sequencing using next generation sequencing (NGS) technologies. The medicinal herbaceous plant, Impatiens balsamina, possess a rich source of diverse bioactive compounds including polysaccharides and phenolic compounds, hindering the extraction of high quality total RNAs. Four existing RNA extraction protocols were performed using fresh leaf material of I. balsamina, namely the CTAB method, the TRI reagent® method, modified TRI reagent® method and a commercial kit but the results were unsatisfactory. This led to the development of a modified CTAB protocol by combining CTAB/ polyvinylpyrrolidone (PVP) extraction with using commercial spin column purification for successful isolation of high quality total RNA from the leaf material of I. balsamina, that is suitable for NGS applications (RIN values > 7). This new method also successfully extracted total RNA from flower buds and capsules of I. balsamina. Penerbit Universiti Kebangsaan Malaysia 2017-06 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/12336/1/46_02_11.pdf Foong, Lian Chee and Siong Hock Ho, Anthony Hock Ho and Lim, Yang Mooi and Tam, Sheh May (2017) A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies. Malaysian Applied Biology, 46 (2). pp. 87-95. ISSN 0126-8643 http://www.mabjournal.com/index.php?option=com_content&view=article&id=644&catid=59:current-view&Itemid=56
repository_type Digital Repository
institution_category Local University
institution Universiti Kebangasaan Malaysia
building UKM Institutional Repository
collection Online Access
language English
description High RNA quality, integrity and reproducibility are essential for functional genomics study involving library preparation and sequencing using next generation sequencing (NGS) technologies. The medicinal herbaceous plant, Impatiens balsamina, possess a rich source of diverse bioactive compounds including polysaccharides and phenolic compounds, hindering the extraction of high quality total RNAs. Four existing RNA extraction protocols were performed using fresh leaf material of I. balsamina, namely the CTAB method, the TRI reagent® method, modified TRI reagent® method and a commercial kit but the results were unsatisfactory. This led to the development of a modified CTAB protocol by combining CTAB/ polyvinylpyrrolidone (PVP) extraction with using commercial spin column purification for successful isolation of high quality total RNA from the leaf material of I. balsamina, that is suitable for NGS applications (RIN values > 7). This new method also successfully extracted total RNA from flower buds and capsules of I. balsamina.
format Article
author Foong, Lian Chee
Siong Hock Ho, Anthony Hock Ho
Lim, Yang Mooi
Tam, Sheh May
spellingShingle Foong, Lian Chee
Siong Hock Ho, Anthony Hock Ho
Lim, Yang Mooi
Tam, Sheh May
A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies
author_facet Foong, Lian Chee
Siong Hock Ho, Anthony Hock Ho
Lim, Yang Mooi
Tam, Sheh May
author_sort Foong, Lian Chee
title A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies
title_short A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies
title_full A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies
title_fullStr A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies
title_full_unstemmed A modified ctab-based protocol for total RNA extraction from the medicinal plant Impatiens balsamina (Balsaminaceae) for next-generation sequencing studies
title_sort modified ctab-based protocol for total rna extraction from the medicinal plant impatiens balsamina (balsaminaceae) for next-generation sequencing studies
publisher Penerbit Universiti Kebangsaan Malaysia
publishDate 2017
url http://journalarticle.ukm.my/12336/
http://journalarticle.ukm.my/12336/
http://journalarticle.ukm.my/12336/1/46_02_11.pdf
first_indexed 2023-09-18T20:02:22Z
last_indexed 2023-09-18T20:02:22Z
_version_ 1777406934216867840

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