title: GSM8877128: GM3_RNA; Mus musculus; RNA-Seq
library_construction_protocol: Sorted cells were centrifuged at 2500 g for 5 mins at 4°C and stored in -80°C. RNA was extracted from FISCs (~200,000 cells per sample) using the RNeasy Plus Micro Kit (Qiagen) according to manufacturer's instructions. Reverse transcription was performed with 1-10 ng RNA using the SMART-Seq v4 Ultra Low Input RNA Kit (Takara) according to manufacturer's instructions. After purification of amplified cDNA, quality and quantity was determined using a 2100 Bioanalyzer the DNA high sensitivity kit (Agilent). RNAseq library constructions were performed using the SMART-Seq Library Prep Kit (Takara) according to manufacturer's instructions. Equimolar amounts of each library were combined, and the pooled library was further quantified using a NEBNext Library Quant Kit (NEB). The RNA-seq libraries were subjected to two rounds of 75 bp paired end sequencing on a NextSeq 550 platform using a 150 cycle kit (Illumina).
identifiers.namespace: GEO
identifiers.id: GSM8877128_r1