GSM9026935: 10_crtx_IgG; Mus musculus; RNA-Seq
For RNA analysis, brain tissues were promptly dissected on ice, flash-frozen by placing on dry ice, and stored at -80°C until further use. RNA isolation was carried out using the RNeasy Micro Kit (Qiagen), following the manufacturer's protocol. RNA quantity and integrity were evaluated using the High Sensitivity RNA assay on the TapeStation 4200 system (Agilent). For RNA sequencing, mRNA libraries were constructed using the TruSeq Stranded mRNA Library Prep Kit (Illumina), with 200 ng of total RNA as the input material. The final libraries were quantified using a Qubit Fluorometer with the Qubit dsDNA HS Assay (Thermo Fisher) and the fragment size distribution was measured using the Agilent high sensitivity D1000 assay on a TapeStation 4200 system (Agilent Technologies). Sequencing was performed in paired-end mode (75 cycles) on a NovaSeq 6000 system (Illumina) using the NovaSeq S1 v1.5 (200 Cycles) chemistry. Raw sequencing data were demultiplexed using bcl2fastq2 v2.20.