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RNA-seq

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Updated: May 9, 2025

RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing

RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing

Published on: August 7, 2021

TIRE-seq simplifies transcriptomics via integrated RNA capture and library preparation.

Piper O'Keeffe^1,2, Yasmin Nouri^1,2, Hui Shi Saw^1,2

¹The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade VIC, Melbourne, VIC, 3052, Australia.

Scientific Reports

|May 2, 2025

Summary

This summary is machine-generated.

Turbocapture Integrated RNA Expression Sequencing (TIRE-seq) streamlines gene expression studies by integrating mRNA purification into library preparation. This method reduces costs and time while improving data quality for large-scale research.

Keywords:

Extraction High throughput RNA Transcriptomics

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Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations

Targeted RNA Sequencing Assay to Characterize Gene Expression and Genomic Alterations

Published on: August 4, 2016

Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

RNA sequencing (RNA-seq) is crucial for understanding gene expression but traditionally involves time-consuming RNA extraction.
Existing methods can lead to sample loss and increased costs in complex research workflows.

Purpose of the Study:

To develop a streamlined RNA sequencing method that eliminates the need for separate RNA extraction.
To improve efficiency, reduce sample loss, and enhance data quality in gene expression analysis.

Main Methods:

Developed TIRE-seq (Turbocapture Integrated RNA Expression Sequencing), integrating mRNA purification into library preparation.
Compared TIRE-seq with the Prime-seq protocol using crude cell lysates.
Applied TIRE-seq to analyze gene expression in stimulated human T cells, differentiating murine dendritic cells, and temozolomide-treated neurospheres.

Main Results:

TIRE-seq demonstrated superior sequencing efficiency compared to Prime-seq with crude cell lysates.
Successfully captured transcriptional changes in activated T cells and identified key genes in dendritic cell differentiation.
Identified differentially expressed genes and pathways in response to temozolomide in patient-derived neurospheres.

Conclusions:

TIRE-seq offers a simplified, cost-effective workflow for gene expression studies.
The method enhances sequencing efficiency and data quality, making it suitable for large-scale research.
TIRE-seq is versatile, applicable to diverse biological systems and research questions.