Feb 08, 2024
Bulk tissue RNA-sequencing - hiPSC-derived astrocytes, neurons and co-cultures
- 1UCL Queen Square Institute of Neurology;
- 2The Francis Crick Institute
Protocol Citation: Karishma D'Sa 2024. Bulk tissue RNA-sequencing - hiPSC-derived astrocytes, neurons and co-cultures . protocols.io https://dx.doi.org/10.17504/protocols.io.ewov1q6kpgr2/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 08, 2024
Last Modified: February 08, 2024
Protocol Integer ID: 94893
Abstract
Bulk tissue RNA-sequencing of hiPSC-derived astrocytes, neurons and co-cultures using the Illumina TruSeq Stranded mRNA Library Prep kit
Bulk tissue RNA-sequencing
Bulk tissue RNA-sequencing
Libraries for sequencing were prepared using the Illumina TruSeq Stranded mRNA Library Prep kit by loading 50 ng of total RNA into the initial reaction; fragmentation and PCR steps were undertaken as per the manufacturer’s instructions.
Final library concentrations were determined using Qubit 2.0 fluorometer and pooled to a normalized input library.
Pools were sequenced using the Illumina NovaSeq 6000 Sequencing system to generate 150 bp paired-end reads with an average read depth of ~137 million paired-end reads per sample.