title: GSM7596352: No_index_frozen_1; Mus musculus; RNA-Seq
library_construction_protocol: Mouse tissues were ground in liquid nitrogen using a tamping tank. Cold tissue powder was rapidly transferred into a 1.5-ml centrifuge tube. Cell line was washed using PBS. Cell pellet (cell line or tissue powder) was lysed in 1 ml ice-cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20, 1% Murine RNase inhibitor (Vazyme Biotech), 0.1% IGEPAL CA-630, 1 mM DTT) and incubated on ice for 2 minutes (tissue), 5 minutes (cell line). Then, 5 ml ice-cold RSBT buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, 0.1% Tween-20) was added to quench the lysis reaction. The medium was then filtered using a 40μm strainer. The nuclei were centrifuged at 500 g for 5 min at 4 ℃ and washed with 1 ml cold RSBT buffer. Library preparations were performed with the HH-seq protocol