GSM7900373: 7day-adECM+ASC+L6; Rattus norvegicus; RNA-Seq
Then the tissue were transferred to cold nuclear lysis buffer (NST with 0.1% NP40, 10 mM Tris-HCl, 146 mM NaCl, 1mM CaCl2, 21 mM MgCl2 and 40 U/mL RNase inhibitor) for 7 min. Add 1 mL ST Wash buffer (10 mM Tris-HCl, 146 mM NaCl, 1 mM CaCl2, 21 mM MgCl2, 0.01% BSA (NEB B9000S) and 40 U/mL RNase inhibitor. Then use 40 um Cell Strainer for collecting nuclear suspension under the 4 ℃, 500 g for 5 min. Then use 100 µL PBS + 1% BSA resuspended the nuclear. After calculating the nuclear number, the nuclear suspension was diluted to 700-1200/µl with PBS + 1% BSA. We used the MobiCube® Single Cell 3' Reagent Kits User Guide (PN-S050400301) to construct the cDNA Library and followed by cell counting, quality control, GEMs generation, barcoding, cDNA amplification and gene expression library construction. The data was submit to DNBSEQ-T7 platform (MGI, China) at PE150 Reads Lengths for final sequencing.