title: GSM7773210: spleen DicerDHEL1het PKRhet MosIR 2211 biol rep 03; Mus musculus; ncRNA-Seq
library_construction_protocol: Organs from adult mice (9-13 w) were homogenized in Qiazol lysis reagent (Qiagen) and total RNA was isolated by phenol–chloroform extraction according to the manufacturer's protocol. Small-RNA libraries were prepared using the NextFlex Small-RNA-seq v3 kit (Amplicon) according to the manufacturer's protocol; 3′ adapter ligation was performed overnight at 20 °C, 15-18 cycles were used for PCR amplification and gel purification was performed for size selection. For gel purification, libraries were separated on a 2.5% agarose gel using 1× lithium borate buffer and visualized with ethidium bromide. The 140-160 bp fraction was cut off the gel and DNA was isolated using the MinElute Gel Extraction Kit (Qiagen). Final libraries were sequenced by 75-nucleotide single-end reading using the Illumina NextSeq500/550 platform