title: GSM3580264: Crush Tsc2 cKO replicate 1A; Mus musculus; RNA-Seq
library_construction_protocol: 3 days post-injury mice were sacrificed. L4 dorsal root ganglia contralateral to injury were isolated, dissociated with papain and collagenase and passed through 70 um filter. Cells were FACS sorted for GFP-positive cells into Clontech lysis buffer with 5% RiboLock Rnase Inhibitor (Thermo Fisher). Library preparation was performed with ~100 cells collected by flow cytometry in 10 ul of Clontech's reaction buffer using the SMARTer Ultra Low RNA kit for Illumina Sequencing (Clontech) per manufacturer's protocol. Three technical replicates of 100 cells each (A-C) were obtained for each biological replicate (1-4). cDNA was amplified for 13 cycles. cDNA was fragmented using a Covaris E220 sonicator using peak incident power 18, duty cycle 20%, cycles/burst 50, time 120 seconds at room temperature. cDNA was blunt ended, had an A base added to the 3'ends, and then had Illumina sequencing adapters ligated to the ends. Ligated fragments were then amplified for 15 cycles using primers incorporating unique index tags. Fragments were sequenced on an Illumina HiSeq-3000 using single reads extending 50 bases.
attributes.tag: GEO Accession
attributes.value: GSM3580264