GSM5518445: Sciatic nerve, single nuclei RNA-seq SN2; Mus musculus; RNA-Seq
For sciatic nerve snRNA-seq libraries, 2 male and 2 female mice were pooled for each experiment. For vagus nerve snRNA-seq libraries, were prepared from a pool of 9 male and 9 female mice. For peroneal and sural nerve snRNA-seq libraries, 5 male and 5 female mice were pooled for each experiment. Upon dissection, the nerves were immediately transferred to digestion media containing 0.3% collagenase IV (Sigma Aldrich C5138), 0.04% hyaluronidase (Sigma Aldrich H3506) and 0.04% DNase I (Sigma Aldrich DN25)  for 10-15 min at 37C. The cells were filtered through 70 um cell strainers and washed twice with cold 5% PBS containing 0.1% BSA, RNaseIn and SuperaseIn. Cells were mechanically dissociated using a Dounce homognizer on ice; media for mechanical dissociation was prepared according to Krishnaswami's protocol87. The nuclei and cellular debris were filtered through 40 um cell strainers and washed with cold 5% PBS containing 0.1% BSA, RNaseIn (Ambion AM2682) and SuperaseIn (Invitrogen AM2694) and the debris was removed from the nuclei by immediate FACS sorting. The purified nuclei were used to prepare single nuclei RNA-seq libraries following manufacturer's instructions (10X Genomics). A total of 11 single nuclei libraries were prepared: 5 libraries from sciatic nerves from Sun1-GFP/+:ActB-Cre/+ mice; 2 libraries from sciatic nerves from Sun1-GFP/+:Mpz-Cre/+ mice; 2 libraries from vagus nerves from Sun1-GFP/+:ActB-Cre/+ mice; 1 library from peroneal nerves dissected from Sun1-GFP/+:ActB-Cre/+ mice; 1 library from sural nerves dissected from Sun1-GFP/+:ActB-Cre/+ mice. 10X genomics single-cell and single-nuclei RNA-seq libraries single-cell and single-nuclei RNA-seq bam files contain reads generated by both instruments HiSeq2500 / NovaSeq6000