GSM8488765: miRNA_LV23B; Felis catus; RNA-Seq
The miRNeasy Mini Kit (Qiagen, Toronto, ON, Canada) was used to isolate total RNA.RNA purity was assessed using the Nanodrop 2000 spectrophotometer. Samples with a 260/280 ratio of >1.9 and RNA integrity number of >5.7 were used for sequencing. A stranded, paired-end poly-A mRNA library was prepared for each sample. Sequencing libraries were generated using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, USA) as per manufacturer's instructions. Briefly, mRNA was isolated, fragmented and primed from total RNA and cDNA synthesis were performed. The cDNA fragments were adapter ligated and PCR amplification was done according to Illumina instructions.  The libraries were sequenced on the NovaSeq 6000 system, SP flowcell 2 x 150 bp (Illumina).