title: GSM4162465: EAE-m3041-IL23Rhet-GP; Mus musculus; RNA-Seq
library_construction_protocol: single cells were sorted into 96-well PCR plate containing dNTP, 3' RT primer, RNase inhibitor, superscript II buffer for cell lysis and storage at -80C. Libraries from isolated single cells were generated based on the Smart-seq2 protocol (Picelli et al., 2014) with the following modifications. In brief, RNA from sorted single cells was used as template for oligo-dT primed reverse transcription with Superscript II Reverse Transcriptase (Thermo Fisher), and the cDNA library was generated by 20 cycle PCR amplification with IS PCR primer using KAPA HiFi HotStart ReadyMix (KAPA Biosystems) followed by Agencourt AMPure XP bead purification as described. Libraries were tagmented using the Nextera XT Library Prep kit (Illumina) with custom barcode adapters (sequences available upon request). Libraries from 1152 cells with unique barcodes were combined and sequenced on a HiSeq2500 in RapidRun mode, with 2x50 nt reads.
attributes.tag: GEO Accession
attributes.value: GSM4162465