Centro de Bioinnovación Antofagasta (CBIA), Facultad de Ciencias del Mar y de Recursos Biológicos, Universidad de Antofagasta, Antofagasta 1270300, Chile.
Programa de Doctorado en Ciencias Biológicas, Mención en Biología Celular y Molecular, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta 1270300, Chile.
Int J Mol Sci. 2024 Sep 6;25(17):9642. doi: 10.3390/ijms25179642.
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 , 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 frozen at -20 °C, obtained from 400 mL of culture with a total of 10 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.
在有害藻华(HAB)样本中,海水中可能含有大量微生物和元素离子。再加上关键赤潮甲藻等的细胞壁的硬度, 这使得 RNA 提取变得非常困难。这些成分干扰 RNA 分离,导致其降解,此外,HAB 的复杂海水特性也可能阻碍有效 RNA 测序和转录组分析的 RNA 分离。在这项研究中,通过应用 Micropestle 系统对冷冻在 -20°C 的细胞沉淀进行 Trizol 方法的改良,建立了一种 RNA 分离技术,该细胞沉淀来自于 400 mL 培养物,总细胞数为 10 个/mL。改良的 Trizol 方案的结果生成了用于转录组测序的优质 RNA 样品,这是通过在 Analyzer Agilent 4150 上进行测量确定的。