GSM4852336: H-ras infected, RNA-Seq, replicate 3; Mus musculus; RNA-Seq
Not-infected (NI), Empty Vector-infected (EV) and H-Ras-infected (Ras) PMK were cross-linked on ice at 0.15 J cm−2 with UV light at 254 nM. Immediately after irradiation, cells were lysed in 1 mL of lysis buffer (50 mM Tris-HCl, pH 7.4; 100 mM NaCl; 1% NP-40; 0.1% SDS; 0.5% sodium deoxycholate). Cell extracts were sonicated using a bioruptor (Digenode) for 10 cycles of 30 seconds, level L at 4ºC, then they were cleared by centrifugation at 10'000 rpm for 10 min at 4°C. RNA was isolated from the extracts by phenol:chloroform extraction. Truseq total RNA-seq protocol with rRNA depletion