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An efficient and effective RNA extraction protocol for ferns.

Jessie A Pelosi1, Ruth Davenport1, W Brad Barbazuk1,2

  • 1Department of Biology University of Florida Gainesville 32611 Florida USA.

Applications in Plant Sciences
|December 4, 2024
PubMed
Summary

We developed a cetyltrimethylammonium bromide (CTAB)-based RNA extraction protocol for ferns. This method yields high-quality RNA, essential for gene expression analysis and advancing fern genomic resources.

Keywords:
RNA extractioncetyltrimetylammonium bromide (CTAB)fernssecondary compounds

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Area of Science:

  • Plant molecular biology
  • Genomics
  • Biotechnology

Background:

  • High-quality RNA extraction is crucial for gene expression analysis.
  • Extracting RNA from ferns is challenging due to cell wall composition and secondary compounds.
  • Genomic resources for ferns are underdeveloped compared to other plant lineages.

Purpose of the Study:

  • To develop an efficient and reliable RNA extraction protocol for diverse fern species and tissues.
  • To overcome the challenges associated with RNA isolation in ferns.
  • To facilitate the development of genomic resources for ferns.

Main Methods:

  • A cetyltrimethylammonium bromide (CTAB)-based RNA extraction protocol was developed.
  • The protocol was tested across a wide phylogenetic range of ferns and various tissue types.
  • Two variations of the protocol were presented: one with precipitation and one without.

Main Results:

  • The CTAB-based protocol consistently yielded high-quality RNA from multiple fern tissues.
  • RNA extracts met high purity standards with OD 260/280 ratios of 1.9-2.1 and OD 260/230 ratios >1.6.
  • RNA integrity numbers (RIN) were consistently above 7, indicating good RNA quality.

Conclusions:

  • The developed protocol is efficient for extracting high-quality RNA from ferns.
  • This method can be applied across diverse fern species and tissue types.
  • The protocol will aid in closing the gap in genomic resource development for ferns.