GSM7963162
Human DRG tissues were procured from National Disease Research Interchange (NDRI). The research application was approved by the NDRI Feasibility Committee (RLUW1 01). Six DRGs between T11 to L5 of three human donors aged from 23- 61 years were used in this study. The dissected DRG tissues from human donors were immediately imbedded in OCT, shipped to the Luo lab on dry ice and stored in -80 ℃ until use. The information of DRGs and dis-identified donors and screening criteria are summarized in the manuscript Supplementary Tables 1 & 2. The hDRGs imbedded in OCT were cryosectioned (Cryostat Leica Cm1950) into 20 μm sec-tions and mounted onto Arcturus PEN Membrane Frame Slides (Applied biosystems, LCM0521). One of every five consecutive sections was collected for laser capture microdissec-tion to avoid repeated dissection of the soma from the same neuron in different sections. The slides were stored in -80 ℃ until further use.On the day of laser capture microdissection, the slides were transferred to the SBDRC laser capture microdissection (LCM) core on dry ice. Before dissection, the section was briefly stained with RNase free Arcturus™ Histogene™ staining solution (Applied biosystems, 12241-05) for better visualization of neuronal soma: 70% cold EtOH for 30s; Histogene™ staining for 10s; 70% cold EtOH for 30s; 95% cold EtOH for 30s; 100% cold EtOH for 30s and air dry for 2min. Then, the slide was put onto laser capture microdissection microscope system (Leica LMD6) for the neuronal soma dissection. The laser was calibrated, and the laser intensity was adjusted to achieve best dissection efficiency. The dissected individual neuronal soma was col-lected in the cap of a 200 μl PCR tube containing 4 μl lysis buffer. The sequencing library was generated following Smart-seq2 workflow. The libraries passing through all quality con-trols were selected for the final sequencing.