title: GSM8577709: Lung, 30ng papain, 3 days, RNA-seq, replicate1; Mus musculus; RNA-Seq
library_construction_protocol: Female mice were used for library construction. Lung ILC2s (Lin−CD25+ST2+ cells) were sorted by flow cytometry (Beckman Coulter) respectively from naïve (as ILC2nav) or papain-treated (50 μg/day for 3 days) mice (as ILC2eff). Construction of the cDNA library and transcriptome sequencing were performed by BGI Genomics, BGI-Shenzhen. Generally, total RNA was treated with DNase I and purified with magnetic beads with Oligo (dT) to isolate mRNA. Treated with the fragmentation buffer, mRNA was fragmented into short fragments, which were the templates of cDNA library. Short fragments were purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. Then, the short fragments are connected with adapters and suitable fragments were selected for the PCR amplification. Agilent 2100 Bioanalyzer and ABI StepOnePlus Real-Time PCR System were used for quantification and qualification of the cDNA library.