title: GSM8701202: JSL1 WT cells. PIC stimulation, replicate B; Homo sapiens; RNA-Seq
library_construction_protocol: Following an 8-hour treatment with either polyIC or IFN-Alpha 2a, cells were collected, and RNA was isolated using TRIzol reagent according to the manufacturer's instructions (Thermo Fisher Scientific, Catalog Number: 15596026). The RNA integrity was assessed using an Agilent Bioanalyzer, ensuring all samples had an RNA Integrity Number greater than 8. RNA samples were quantified using Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked using Agilent TapeStation 4200 (Agilent Technologies, Palo Alto, CA, USA). RNA sequencing libraries were prepared using the NEBNext Ultra II RNA Library Prep Kit for Illumina using manufacturer's instructions (NEB, Ipswich, MA, USA). Briefly, mRNAs were initially enriched with Oligod(T) beads. Enriched mRNAs were fragmented for 15 minutes at 94 °C. Converted the RNA molecules into complementary DNA (cDNA). cDNA fragments were end repaired and adenylated at 3'ends, and universal adapters were ligated to cDNA fragments, followed by index addition and library enrichment by PCR with limited cycles. The sequencing library was validated on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA) as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA). The sequencing libraries were clustered on a flowcell. After clustering, the flowcell was loaded on the Illumina NovaSeq instrument according to manufacturer's instructions. The samples were sequenced using a 2x150bp Paired End (PE) configuration. Image analysis and base calling were conducted by the Control software. Raw sequence data (.bcl files) generated by the sequencer were converted into fastq files and de-multiplexed using Illumina's bcl2fastq 2.17 software. One mismatch was allowed for index sequence identification.
identifiers.namespace: GEO
identifiers.id: GSM8701202_r1