IL17B treatment 1
Total RNA isolation was performed using TRI Reagent from the Molecular Research Center. Subsequently, the isolation of mRNA was carried out using magnetic beads attached to poly(T) oligonucleotides. For the initiation of first-strand cDNA synthesis, random hexamer primers along with Moloney murine leukemia virus reverse transcriptase (RNase H) were used. The subsequent step involved the preparation of second-strand cDNA using DNA polymerase I and RNase H. Following an end repair process and 3 adenylation, adaptors were ligated to the cDNA fragments. Purification of library fragments was conducted using the AMPure XP system (Beckman Coulter, Brea, CA). Subsequent PCR amplification was performed using Phusion high-fidelity DNA polymerase, universal PCR primers, and Index (X) Primer from NEB (Ipswich, MA, USA). The resulting PCR products were purified using the AMPure XP system, and library quality was assessed using the Agilent Bioanalyzer 2100 system (Agilent, Santa Clara, CA). For cluster generation, the cBot cluster generation system along with the TruSeq PE cluster kit v3-cBot-HS from Illumina (San Diego, CA) was used. The subsequent sequencing process was carried out on a NovaSeq 6000 platform (Illumina) using paired-end reads of 150 bp
SUB15240850
IL17BT1