title: GSM3834144: RNA_ILC2_IL33_CGRP_4h_rep2; Mus musculus; RNA-Seq
library_construction_protocol: FastATAC-seq: 1 X 10^4 cells were pelleted and washed with 50 μl 1× PBS. After pelleting the nuclei by centrifuging at 500 × g for 10 min, the pellets were re-suspended in 50 μl transposase mixture (25 µl of 2x TD buffer, 2.5 µl of TDE1, 0.5 µl of 1% digitonin, 22 µl of nuclease-free water) (#FC-121-1030, Illumina; #G9441, Promega). The reaction was incubated at 37°C with shaking at 300 rpm for 30 min. The fragmentalized DNAs were then purified using a QIAGEN MinElute kit and amplified with 10 or 11 cycles of PCR based on the amplification curve, and the paired-end libraries were purified using a QIAGEN PCR cleanup kit scRNA-seq: Freshly sorted ILCs and Th cells (13,000 cells each) were encapsulated into droplets, and libraries were prepared using Chromium Single Cell 3' Reagent Kits v2 according to manufacturer's protocol (10X Genomics). RNA-seq: Total RNA was extracted from 2-5 x 10^4 cells by Direct-zol™ RNA MicroPrep (Zymo Research). Single-end libraries were prepared from 0.1-0.5 μg of total RNA using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, E7530S)
attributes.tag: GEO Accession
attributes.value: GSM3834144