title: GSM5830148: BulkRNAseq_RANKL_rep_1; Mus musculus; RNA-Seq
library_construction_protocol: Pellets were snap frozen and stored at ‐80 ⁰C. RNA from snap frozen pellets of drug treated organoids was extracted using the RNeasy RNA extraction kit (Qiagen) with DNaseI treatment. Total RNA (1 µg per replicate) was used for library preparations using the KAPA RiboErase Kit (HMR) (Roche) following manufacturer's protocol. A fragmentation step was carried out for 6.5 minutes at 94°C and 7 μM NextFlex DNA barcodes (PerkinElmer) were used for adapter ligation. Libraries were amplified using 6 amplification cycles. Library concentration was measured using the dsDNA Fluorescence Quantification Assays (DeNovix), and library size was determined using the BioAnalyzer High Sensitivity DNA Kit (Agilent Technologies) in a BioAnalyzer instrument (Agilent). Sequencing was performed using an Illumina NextSeq 500, and 42-bp paired-end reads were generated.