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A Modified Protocol for Total Rna Isolation from Different Oil Palm (Elaeis guineensis) Tissues using Cetyltrimethylammonium Bromide


Affiliations
1 Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
2 Department of Environmental Sciences, Lahore College for Women University Lahore, Lahore 54600, Pakistan
3 Institute of Horticultural Sciences, University of Agriculture, Faisalabad, Pakistan
 

Extraction of high-quality RNA from oil palm tissues is challenging due to the presence of polysaccharides, polyphenols and other complexes that co-precipitate with RNA. Therefore, isolation of high-quality RNA from oil palm is challenging due to the presence of varying amounts of these constituents in diverse tissues. This communication describes a modified RNA extraction protocol based on the cetyltrimethylammonium bromide (CTAB) method which is useful for extracting high-quality RNA from different oil palm tissues. Total RNA isolation using a modified CTAB protocol was compared with two different methods, a conventional TRIzol method and the method for RNA isolation from palms (MRIP). Both methods were useful for isolating RNA from leaf tissues; however, they were not effective in isolating RNA from other tissues. The current protocol based on a modified CTAB method was efficient in isolating high-quality total RNA from oil palm fruit tissues, including mesocarp and endosperm, stem, ischolar_main and flower tissues. This modified CTAB-based protocol gave approximately 20-30 μg of total RNA from 150 mg of tissue within 5-6 h.

Keywords

CTAB Method, Gene Expression, Oil Palm, RNA Extraction.
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  • A Modified Protocol for Total Rna Isolation from Different Oil Palm (Elaeis guineensis) Tissues using Cetyltrimethylammonium Bromide

Abstract Views: 458  |  PDF Views: 126

Authors

Rashad Qadri
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Amjad Iqbal
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Yi Wu
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Jing Li
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China
Numrah Nisar
Department of Environmental Sciences, Lahore College for Women University Lahore, Lahore 54600, Pakistan
Muhammad Azam
Institute of Horticultural Sciences, University of Agriculture, Faisalabad, Pakistan
Yaodong Yang
Hainan Key Laaboratory of Tropical Oil Crops Biology/Coconut Research Institute, Chinese Academy of Tropical Agricultural Science, Wenchang, Hainan 571339, China

Abstract


Extraction of high-quality RNA from oil palm tissues is challenging due to the presence of polysaccharides, polyphenols and other complexes that co-precipitate with RNA. Therefore, isolation of high-quality RNA from oil palm is challenging due to the presence of varying amounts of these constituents in diverse tissues. This communication describes a modified RNA extraction protocol based on the cetyltrimethylammonium bromide (CTAB) method which is useful for extracting high-quality RNA from different oil palm tissues. Total RNA isolation using a modified CTAB protocol was compared with two different methods, a conventional TRIzol method and the method for RNA isolation from palms (MRIP). Both methods were useful for isolating RNA from leaf tissues; however, they were not effective in isolating RNA from other tissues. The current protocol based on a modified CTAB method was efficient in isolating high-quality total RNA from oil palm fruit tissues, including mesocarp and endosperm, stem, ischolar_main and flower tissues. This modified CTAB-based protocol gave approximately 20-30 μg of total RNA from 150 mg of tissue within 5-6 h.

Keywords


CTAB Method, Gene Expression, Oil Palm, RNA Extraction.

References





DOI: https://doi.org/10.18520/cs%2Fv116%2Fi3%2F479-482