Improved CTAB method for RNA extraction of thick waxy leaf tissues from sago palm (Metroxylon sagu Rottb.)
Abstract Background There is a growing interest in transcriptomics studies parallel to the advancement of transcriptome databases and bioinformatics, which provided the opportunity to study responses to growths, stimuli and stresses. There is an increase in demand for excellent RNA extraction techni...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2022-09-01
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| Series: | Chemical and Biological Technologies in Agriculture |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40538-022-00329-9 |
| _version_ | 1828148170281975808 |
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| author | Wei-Jie Yan Fifi Hafizzah Pendi Hasnain Hussain |
| author_facet | Wei-Jie Yan Fifi Hafizzah Pendi Hasnain Hussain |
| author_sort | Wei-Jie Yan |
| collection | DOAJ |
| description | Abstract Background There is a growing interest in transcriptomics studies parallel to the advancement of transcriptome databases and bioinformatics, which provided the opportunity to study responses to growths, stimuli and stresses. There is an increase in demand for excellent RNA extraction techniques. General RNA extraction protocols can be used in RNA extraction, but the quality and quantity vary in different types of tissues from different organisms. Hence, a specific RNA extraction method for each organism’s tissue type is required to obtain the desired RNA quality and quantity. Results The improved CTAB RNA extraction method is superior to the PCI method and MRIP method for thick waxy leaves that were applied for mature sago palm (Metroxylon sagu Rottb.) leaf tissue and produce total RNA extract with good purity (OD 260/280 ≥ 1.8, OD 260/230 ≥ 2.0) and integrity (RIN ~ 7). RNA sequencing was conducted with the extracted samples and showed good assembly results (Q20 ≥ 97, Q30 ≥ 91%, assembly mean length ≥ 700 bp). Conclusion The improved CTAB RNA extraction method enables rapid, cost-effective, and relatively simple RNA extraction from waxy, fibrous and high-in-polyphenol sago palm (M. sagu Rottb.) leaf tissue with next-generation RNA sequencing recommended quality. Graphical Abstract |
| first_indexed | 2024-04-11T21:12:06Z |
| format | Article |
| id | doaj.art-9a923e6f43bd4d2da888608bd0636235 |
| institution | Directory Open Access Journal |
| issn | 2196-5641 |
| language | English |
| last_indexed | 2024-04-11T21:12:06Z |
| publishDate | 2022-09-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Chemical and Biological Technologies in Agriculture |
| spelling | doaj.art-9a923e6f43bd4d2da888608bd06362352022-12-22T04:03:00ZengSpringerOpenChemical and Biological Technologies in Agriculture2196-56412022-09-019111410.1186/s40538-022-00329-9Improved CTAB method for RNA extraction of thick waxy leaf tissues from sago palm (Metroxylon sagu Rottb.)Wei-Jie Yan0Fifi Hafizzah Pendi1Hasnain Hussain2Centre for Sago Research, Faculty of Resource Science and Technology, Universiti Malaysia SarawakCentre for Sago Research, Faculty of Resource Science and Technology, Universiti Malaysia SarawakCentre for Sago Research, Faculty of Resource Science and Technology, Universiti Malaysia SarawakAbstract Background There is a growing interest in transcriptomics studies parallel to the advancement of transcriptome databases and bioinformatics, which provided the opportunity to study responses to growths, stimuli and stresses. There is an increase in demand for excellent RNA extraction techniques. General RNA extraction protocols can be used in RNA extraction, but the quality and quantity vary in different types of tissues from different organisms. Hence, a specific RNA extraction method for each organism’s tissue type is required to obtain the desired RNA quality and quantity. Results The improved CTAB RNA extraction method is superior to the PCI method and MRIP method for thick waxy leaves that were applied for mature sago palm (Metroxylon sagu Rottb.) leaf tissue and produce total RNA extract with good purity (OD 260/280 ≥ 1.8, OD 260/230 ≥ 2.0) and integrity (RIN ~ 7). RNA sequencing was conducted with the extracted samples and showed good assembly results (Q20 ≥ 97, Q30 ≥ 91%, assembly mean length ≥ 700 bp). Conclusion The improved CTAB RNA extraction method enables rapid, cost-effective, and relatively simple RNA extraction from waxy, fibrous and high-in-polyphenol sago palm (M. sagu Rottb.) leaf tissue with next-generation RNA sequencing recommended quality. Graphical Abstracthttps://doi.org/10.1186/s40538-022-00329-9Metroxylon saguRNA extractionTranscriptomicsImproved CTAB method |
| spellingShingle | Wei-Jie Yan Fifi Hafizzah Pendi Hasnain Hussain Improved CTAB method for RNA extraction of thick waxy leaf tissues from sago palm (Metroxylon sagu Rottb.) Chemical and Biological Technologies in Agriculture Metroxylon sagu RNA extraction Transcriptomics Improved CTAB method |
| title | Improved CTAB method for RNA extraction of thick waxy leaf tissues from sago palm (Metroxylon sagu Rottb.) |
| title_full | Improved CTAB method for RNA extraction of thick waxy leaf tissues from sago palm (Metroxylon sagu Rottb.) |
| title_fullStr | Improved CTAB method for RNA extraction of thick waxy leaf tissues from sago palm (Metroxylon sagu Rottb.) |
| title_full_unstemmed | Improved CTAB method for RNA extraction of thick waxy leaf tissues from sago palm (Metroxylon sagu Rottb.) |
| title_short | Improved CTAB method for RNA extraction of thick waxy leaf tissues from sago palm (Metroxylon sagu Rottb.) |
| title_sort | improved ctab method for rna extraction of thick waxy leaf tissues from sago palm metroxylon sagu rottb |
| topic | Metroxylon sagu RNA extraction Transcriptomics Improved CTAB method |
| url | https://doi.org/10.1186/s40538-022-00329-9 |
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