GSM2176985: ILC210 08-10Bit22IL10pos; Mus musculus; RNA-Seq
RNA was extracted and purified from lung (ILC2) or spleen (Treg) using the RNeasy Plus Mini Kit (Qiagen) per manufacturers instructions Libraries of ILC2PBS and ILC2IL-33 samples were constructed using SMARTer Ultra Low Input RNA v3 kit (Clontech) as previously described. Briefly, the protocols for enzymatic fragmentation and ligation of the Ion Xpress Plus Fragment Library Kit (Life Technologies) were adjusted for our low-input total RNA of samples. Then resulting cDNA libraries were amplified onto Ion Sphere Particles with an Ion PI template OT2 200 Kit v3 (Life Technologies) and sequenced on an Ion Proton (Life Technologies) to generate an average depth of 20 million single-end 200bp reads, with less than 6.7% of the reads from ribosomal RNA and over 94% of the reads mapping to the mouse genome. Similarly, libraries of ILC2_act, ILC2_10, Treg_10- and Treg_10+ samples were constructed using SMARTer Ultra Low Input RNA v3 kit (Clontech), indexed using Nextera indices (Illumina) and sequenced on a NextSeq 500 (Illumina) using 75bp single-end sequencing kit per the manufacturer’s instructions. On average, about 10 million reads were generated from each sample.
GEO Accession
GSM2176985