title: GSM9117772: 4h_CNT_F_S_S2; Mus musculus; miRNA-Seq
library_construction_protocol: The two trigeminal ganglia and two superior cervical ganglia were removed surgically from each mouse. Ganglia were homogenized in 700 µL QIAzol using a pellet pestle motor and RNA was extracted using the miRNeasy Mini Kit (Qiagen, 217004) according to the manufacturer's protocol, followed by RNA concentration determination (NanoDrop 2000, Thermo Scientific). Average trigeminal ganglia yield was 193 ng/µL and average superior cervical ganglia yield was 64 ng/µL. RIN was determined for all samples (Bioanalyzer 6000, Agilent), with an average of 8.1 (range 5.3-9.3). Half of the samples (48 out of 96, see Table 1) were selected for short RNA-seq, 2 out of the 4 in each subgroup above. Average RIN for these 48 samples was 8.3 (range 6.4-9.3). Libraries were constructed from 140 ng total RNA of each sample, using the NEBNext Multiplex Small RNA library prep kit for Illumina (New England Biolabs, NEB-E7560S) and the small RNA fraction was sequenced on the NextSeq 500 System (Illumina) at the Center for Genomic Technologies Facility, the Hebrew University of Jerusalem.