GSM1446368: Replicate 2, RNA-Seq, LPS-stimulus, time 1h; Mus musculus; RNA-Seq
RNA was extracted from cells using RNeasy Mini Kit (Quiagen), according to the manufacturer’s protocol. Enrichment of polyadenylated RNA (polyA+ RNA) from total RNA was performed using Oligo(dT) dynabeads (Invitrogen) according to the manufacturer’s protocol. The mRNA was chemically fragmented into ~80-nt-long fragments using RNA fragmentation reagent (Ambion), followed by Turbo DNase treatment (Ambion). Strand-specific RNA-seq libraries were generated as previously described (Engreitz et al., Science, 2013). Briefly, RNA was first subjected to FastAP Thermosensitive Alkaline Phosphatase (Thermo Scientific), followed by a 3’ ligation of an RNA adapter using T4 ligase (New England Biolabs). Ligated RNA was reverse transcribed using AffinityScript Multiple Temperature Reverse Transcriptase (Agilent), and the cDNA was subjected to a 3’ ligation with a second adapter using T4 ligase. The single-stranded cDNA product was then amplified for 9- to 14 cycles in a PCR reaction. Libraries were sequenced on an Illumina HiSeq 2500 generating 30bp paired-end reads.
GEO Accession
GSM1446368