Malays. Appl. Biol. (2017) 46(2): 87-95
A MODIFIED CTAB-BASED PROTOCOL FOR TOTAL RNA
EXTRACTION FROM THE MEDICINAL PLANT
Impatiens balsamina (BALSAMINACEAE) FOR
NEXT-GENERATION SEQUENCING STUDIES
LIAN CHEE FOONG1, ANTHONY SIONG HOCK HO1, YANG MOOI LIM2
and SHEH MAY TAM1*
1School of Biosciences, Taylor’s University, Jalan Taylors, 47500 Subang Jaya,
Selangor, Malaysia
2Department of Pre-Clinical Sciences, Faculty of Medicine and Health Sciences,
Universiti Tunku Abdul Rahman, Lot PT 21144, Jalan Sungai Long,
Bandar Sungai Long, 43000 Kajang, Selangor, Malaysia.
*Email: This e-mail address is being protected from spambots. You need JavaScript enabled to view it
Accepted 20 April 2017, Published online 27 June 2017
ABSTRACT
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.
Key words: Impatiens balsamina, total RNA extraction, CTAB protocol
