Abstract
Isolation of high-quality RNA free of contaminants, such as polyphenols, proteins, plant secondary metabolites, and genomic DNA from plant tissues, is usually a challenging but crucial step for molecular analysis. We developed a novel protocol based on the cetyltrimethylammonium bromide method to isolate high-quality RNA from blackberry plant tissues, especially fruits. Most DNA was removed when acetic acid was utilized, before RNA precipitation. Thus, lithium chloride, a reagent widely used for RNA purification, was not needed. The isolation time was shortened to less than 3 h. The RNA was quite pure, with little DNA contamination. The quality of the RNA was assessed by spectrophotometric readings and electrophoresis on agarose gels. It was good enough for downstream enzymatic reactions, such as reverse transcription-PCR, cloning and real-time PCR assay. The method yielded an amount of total RNA comparable to previously described protocols.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.