title: GSM9086571: Het-E-4_S1203; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was purified from 2-5 x 10^5 cells with TRIzol (Invitrogen) and 1-2 mg RNA was treated with DNase I (Thermo Fisher Scientific) for 15 min at room temperature. After heat inactivation of DNase I with EDTA for 10 min at 65°C, 0.5-1 mg RNA was incubated with a 4:1 mixture of oligo(dT) primers and random hexamer at 68°C for 10 min. RNA/primers were incubated with Moloney murine leukemia virus reverse transcription (M-MLV RT) (Thermo Fisher Scientific), 5X first strand buffer (Thermo Fisher Scientific), 10 mM dithiothreitol (Thermo Fisher Scientific), RNAsin (Promega), and 0.5 mM deoxynucleoside triphosphates (New England Biolabs) at 42°C for 1 h and then heat inactivated at 95°C for 5 min. RNA library was prepared using TruSeq Stranded Total RNA (rRNA depletion) Library kit following manufacturer's protocol.