RNAseq Kiss1 Neurons

Pools of 10 - 15 Kiss1 neurons from the rostral (R samples) or caudal (C samples) regions of the arcuate or the RP3V region (A samples) were transferred to a separate PCR tube containing a 8 μl of water, 1 μl 0.1mM DTT and 1 μl RNase inhibitor (40U RNase OUT, Invitrogen) and snap-frozen on dry ice and kept at -80⸰C until further analysis. Library preparation was performed using SMART-Seq v4 Ultra Low Input RNA Kit (Takara Bio USA, Inc. Cat#: 634889). Samples were sequenced induplicate using single-end 75 bp rreads on a NextSeq500 (Illumina, USA) machine using NextSeq 500/550 High Output v2 Kit (75 cycles) (Illumina, USA). The sequencing depth of the raw reads ranged from 23-49 million per sample.