Nova6000 plate high output model

The total RNA (1.0 μg/sample) was subjected to rRNA depletion by Ribo-Zero rRNA Removal (Human/Mouse/Rat) Kit (Illumina, San Diego, CA, United States). Strand-specific RNA libraries were prepared using TruSeq Stranded Total RNA Sample Preparation Kit (Illumina) without poly-A selection. All assays were performed according to the manufacturer’s instructions. RNA-seq was performed on the Illumina Nova6000 plate High Output Model (Illumina, San Diego, United States) (Hrdlickova et al., 2017 (link)), in a 2 × 150-bp paired-end configuration, with a total of more than 2,666 M reads per lane (at least 50 M reads per sample).

Three biological replicates (three mice) from each age group were used in the RNA-seq analysis. Total RNA extracted above was subjected to rRNA depletion by Ribo-Zero rRNA Removal (Human/Mouse/Rat) Kit (Illumina, San Diego, CA, United States). Strand-specific RNA libraries were prepared using TruSeq Stranded Total RNA Sample Preparation Kit (Illumina) without poly-A selection. All assays were performed according to the manufacturer’s instructions. Sequencing was carried out at the Illumina Nova6000 plate High Output Model (Illumina, San Diego, CA, United States) (Hrdlickova et al., 2017 (link)), in a 2 × 150 bp paired-end configuration, with a total of more than 2,666 M reads per lane (at least 40 M reads per sample). All sequencing data are available in NCBI database (accession number: SRP271007).

The extracted total RNA was processed for rRNA depletion using the Ribo-Zero rRNA Removal (Human/Mouse/Rat) Kit (Illumina, San Diego, CA, United States). Using the TruSeq Stranded Total RNA Sample Preparation Kit (Illumina), strand-specific RNA libraries were generated without poly-A selection, according to the manufacturer’s guidelines. Then, RNA-seq was performed on the Illumina Nova 6,000 plate High Output Model (Illumina, San Diego, United States) with over 1,087 M reads per lane.