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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...

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Updated: Aug 18, 2025

Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing

Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing

Published on: October 12, 2018

Large-Scale Single-Nucleus RNA Sequencing Compatible with Complex Archived Samples.

Chao-Yu Liu¹, Hsu-Hsin Chen²

¹College of Medicine, National Taiwan University, Taipei, Taiwan.

Methods in Molecular Biology (Clifton, N.J.)

|December 9, 2022

Summary

This summary is machine-generated.

This study presents a single-nucleus RNA sequencing protocol for frozen ocular tissues, enabling detailed analysis of cell types in conditions like retinitis pigmentosa.

Keywords:

Deep sequencing Gene expression profiling Parallel mRNA analysis Single-cell transcriptomics sNuc-seq

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Nuclei Isolation from Fresh Frozen Brain Tumors for Single-Nucleus RNA-seq and ATAC-seq

Nuclei Isolation from Fresh Frozen Brain Tumors for Single-Nucleus RNA-seq and ATAC-seq

Published on: August 25, 2020

Area of Science:

Ophthalmology
Genomics
Molecular Biology

Background:

Single-cell resolution transcriptome profiling is crucial for understanding complex tissues.
Retinitis pigmentosa involves degenerating ocular tissues with high cellular heterogeneity.

Purpose of the Study:

To introduce a medium-throughput single-nucleus RNA sequencing (snRNA-seq) protocol.
To enable functional cell type and state assessment in frozen ocular tissues.

Main Methods:

Detailed protocol for snRNA-seq using frozen tissue.
Medium-throughput approach suitable for researchers with basic molecular biology training.
Efficient sample processing: ~20,000 nuclei per eye from 3-4 library preparations.

Main Results:

The protocol allows for the identification of functional cell types and states.
Applicable to degenerating ocular tissues, such as those found in retinitis pigmentosa.
Routine yield of approximately 20,000 nuclei per eye is achievable.

Conclusions:

This snRNA-seq protocol provides a valuable tool for studying ocular tissue heterogeneity.
Facilitates research into cell-specific changes in retinal degenerative diseases.
Accessible methodology for broad research application.