GSM4566250: tg_APCs3; Mus musculus; RNA-Seq
Total RNA from freshly FACS-sorted cells was extracted and purified using the RNAqueous Micro total RNA isolation kit (ThermoFisher Scientific, #AM1931). RNA samples were quantified using Qubit 2.0 Fluorometer (Life Technologies, Carlsbad, CA, USA) and RNA integrity was checked with Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA). Starting from total RNA, the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) is used to isolate pure mRNA. Enriched mRNAs were fragmented for 15 minutes at 94 °C. First strand and second strand cDNA were subsequently synthesized. With NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB #E7770), cDNA fragments were end repaired and adenylated at 3'ends, and universal adapter was ligated to cDNA fragments, followed by index addition and library enrichment with limited cycle PCR. The sequencing library was validated on the Agilent TapeStation (Agilent Technologies, Palo Alto, CA, USA), and quantified by using Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA).
GEO Accession
GSM4566250