从人诱导多能干细胞来源的视网膜类器官中分离 RNA。

RNA Isolation from Human Stem Cell-Derived Retinal Organoids.

机构信息

Department of Ophthalmology | Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.

出版信息

Methods Mol Biol. 2024;2822:3-11. doi: 10.1007/978-1-0716-3918-4_1.

Abstract

RNA isolation is an essential first step for many types of molecular analyses, including reverse transcription PCR (RT-PCR)/quantitative RT-PCR (qRT-PCR), Northern blotting, microarrays, and RNA-sequencing. While many RNA purification methods have been reported, it can be challenging to extract sufficient quantity, and suitable quality, of RNA from very small amounts of tissue and/or samples containing low numbers of cells. Here we outline a total RNA isolation method that reproducibly yields high-quality RNA from human stem cell-derived retinal organoids for downstream transcriptomic analysis.

摘要

RNA 分离是许多类型的分子分析的第一步，包括逆转录 PCR（RT-PCR）/定量 RT-PCR（qRT-PCR）、Northern 印迹、微阵列和 RNA 测序。虽然已经报道了许多 RNA 纯化方法，但从非常少量的组织和/或含有少量细胞的样品中提取足够数量和合适质量的 RNA 可能具有挑战性。在这里，我们概述了一种总 RNA 分离方法，该方法可从人诱导多能干细胞衍生的视网膜类器官中重复性地获得高质量 RNA，用于下游转录组分析。