title: GSM5047944: HD-sRNAs-CB; Homo sapiens; miRNA-Seq
library_construction_protocol: small RNA (sRNA) fractions were purified from total RNA with the RNA Clean & Concentrator-5 Kit (Zymo Research; R1015) according to the manufacturer's instructions. CTL-sRNA-PT, HD-sRNA-PT, CTL-sRNA-CTX, HD-sRNA-CTX, CTL-sRNA-CB and HD-sRNA-CB pools (200 ng) were processed with NEBNext® Small RNA Library Prep Set for Illumina® (New England Biolabs; E7330S), size selection was performed by Novex® Pre-Cast 6% TBE PAGE gel (Thermo Fisher Scientific; EC62652BOX) and the bands of interest cut as follows: a) 145-160 bp band (corresponding to adapter-ligated constructs derived from the 21 and 30 nt RNA fragments, respectively), b) 160-180 bp band, c) 180-200 bp band, d) 200-300 bp band, e) 300-500 bp. Paired-end sequencing (2 × 50 cycles, CTL-sRNA-PT, HD-sRNA-PT) or single-end sequencing (1 x 50 nt, CTL-sRNA-CTX, HD-sRNA-CTX, CTL-sRNA-CB, HD-sRNA-CB) of indexed cDNA libraries were then carried out on a HiSeq 2500 machine (Illumina), generating at least 50 million reads per sample. Sequencing was performed using v4 SBS and Cluster Kits (Illumina; FC-401-4002).