Nuclei isolation methods for single-nucleus sequencing

Single-nucleus RNA sequencing (snRNA-seq) is a complementary technology to single-cell RNA sequencing (scRNA-seq) which utilizes isolated nuclei instead of intact cells to profile gene expression. Both technologies became powerful tools for investigating cell types, dynamic states, and functional processes at the single-cell level, profoundly revolutionizing transcriptomic studies. While for scRNA-seq the input material is a freshly isolated tissue, with snRNA-seq isolated nuclei from frozen or preserved tissue can be utilized, enabling processing of archived samples. Furthermore, snRNA-seq can generate  gene expression profile in cells that are difficult to isolate (e.g. adipocytes, neurons, hepatocytes) or whom size exceeds the  technical capabilities of the single-cell technologies.

To achieve a successful snRNA-seq experiment, a key prerequisite is preparing a high-quality a single-nucleus suspension. Isolating good quality nuclei can be a limiting step when working with fragile, archived tissues of variable quality. Furthermore, each nuclei isolation needs to be tailored on the tissue type, state and complexity, a “do it all” protocol being hardly possible. With this two days course we intend to make participants familiar with good-practice-guidelines and methods for nuclei isolation for single-nucleus sequencing with practical sessions included. The course will cover both theoretical and hands-on sessions on sample preparation, tissue lysis, clean-up techniques, quality control and data analysis.