GSM5766485: G795-06 6 P; Mus musculus; RNA-Seq
GSM5766485
RNA extraction was performed using the Picopure RNA Isolation Kit (Thermo Fisher Scientific). RNAs were amplified by two rounds of in vitro transcription using the Arcturus RiboAmp HS PLUS Kit (Thermo Fisher Scientific) according to the manufacturer's protocols. The cDNA libraries were prepared using the KAPA RNA HyperPrep Kit (Roche) as per the manufacturer's instructions. The unique dual barcode sequences were incorporated in the adaptors for multiplexed high-throughput sequencing (NEXTFLEX® Unique Dual Index Barcodes, BioO Scientific). The final product was assessed for its size distribution and concentration using BioAnalyzer High Sensitivity DNA Kit (Agilent Technologies). The libraries were pooled and diluted to 3 nM using 10 mM Tris-HCl, pH 8.5 and then denatured using the Illumina protocol. The denatured libraries were loaded onto an S1 flow cell on an Illumina NovaSeq 6000 (Illumina) and run for 57-65 cycles according to the manufacturer's instructions.