Bulk tissue RNA-sequencing - hiPSC-derived astrocytes, neurons and co-cultures
Karishma D'Sa
Published: 2024-02-09 DOI: 10.17504/protocols.io.ewov1q6kpgr2/v1
Abstract
Bulk tissue RNA-sequencing of hiPSC-derived astrocytes, neurons and co-cultures using the Illumina TruSeq Stranded mRNA Library Prep kit
Steps
Bulk tissue RNA-sequencing
1.
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.
2.
Final library concentrations were determined using Qubit 2.0 fluorometer and pooled to a normalized input library.
3.
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.
