RESUMO

In samples of harmful algal blooms (HABs), seawater can contain a high abundance of microorganisms and elemental ions. Along with the hardness of the walls of key HAB dinoflagellates such as Prorocentrum triestinum, this makes RNA extraction very difficult. These components interfere with RNA isolation, causing its degradation, in addition to the complex seawater properties of HABs that could hinder RNA isolation for effective RNA sequencing and transcriptome profiling. In this study, an RNA isolation technique was established through the modification of the Trizol method by applying the Micropestle System on cell pellets of P. triestinum frozen at -20 °C, obtained from 400 mL of culture with a total of 107 cells/mL. The results of the modified Trizol protocol generated quality RNA samples for transcriptomics sequencing, as determined by their measurement in Analyzer Agilent 4150.

Assuntos

Dinoflagellida , Dinoflagellida/genética , RNA/isolamento & purificação , RNA/genética , Guanidinas/química , Análise de Sequência de RNA/métodos , Proliferação Nociva de Algas , Perfilação da Expressão Gênica/métodos , Transcriptoma , Nucleotídeos/genética , Nucleotídeos/isolamento & purificação , Água do Mar , Fenóis