Unité de Biologie des Spirochètes, Institut Pasteur, Paris, France.
Methods Mol Biol. 2020;2134:41-51. doi: 10.1007/978-1-0716-0459-5_5.
Establishing a rapid method to obtain pure and intact RNA molecules has revolutionized the field of RNA biology, enabling laboratories to routinely perform RNA analysis such as Northern blot, reverse transcriptase quantitative PCR, and RNA sequencing. Here, we describe an application of the effective single-step method of RNA extraction (or guanidinium thiocyanate-phenol-chloroform extraction) applied to Leptospira species. This method is based on the powerful ability of guanidinium thiocyanate to inactivate RNases and on the different solubilities of RNA and DNA in acidic phenol. This method allows one to reproducibly obtain total RNAs with high yield and integrity, as determined by capillary electrophoresis, suitable for the RNA sequencing technology.
建立一种快速获得纯净完整 RNA 分子的方法彻底改变了 RNA 生物学领域，使实验室能够常规进行 RNA 分析，如 Northern blot、逆转录定量 PCR 和 RNA 测序。在这里，我们描述了有效一步法 RNA 提取（或异硫氰酸胍-酚-氯仿提取）在钩端螺旋体属中的应用。该方法基于异硫氰酸胍能够使 RNase 失活的强大能力，以及 RNA 和 DNA 在酸性酚中的不同溶解度。该方法能够重复获得高产量和完整性的总 RNA，这可以通过毛细管电泳确定，适用于 RNA 测序技术。
