title: GSM8371690: GFPactiveTLR4siRNAHC_31; Mus musculus; RNA-Seq
library_construction_protocol: For the isolation of total RNA from cells isolated by fluorescence- activated cell sorting, the RNeasy Plus Micro Kit (Qiagen, 74034), according to manufacturer's protocol. 3'mRNA sequencing libraries were prepared using Lexogen QuantSeq 3'mRNA-Seq v2 Library Prep Kit FWD with UDIs and the UMI Second Strand Synthesis Module  following the manufacturer's protocol. Prior to library preparation, the concentration of RNA was measured using the Promega Quantus Fluorometer. Additionally, the size distribution of the RNA was assessed using the Agilent TapeStation with an RNA ScreenTape. Quantifiaction and quality assessment were repeated after library preparation, again using the Quantus fluorometer and the Agilent TapeStation with a High Sensitivity D1000 ScreenTape. Libraries were denatured, diluted, and loaded onto a NextSeq High Output v2.5 (75 cycles) flow cell. The 20% PhiX control library was spiked in to improve base calling accuracy. Single-end sequencing was performed with 75 cycles on the Illumina NextSeq platform according to the manufacturer's instructions.