GSM3350423: Tg2a_2i_3; Mus musculus; RNA-Seq
RNA: Cells were lysed with 1mL TRIzol (ThermoFisher) and RNAs extracted according to manufacturer's protocol. To eliminate any genomic DNA contamination, this was followed by an additional DNAse I treatment (Qiagen 79254) for 20min at 37°C followed by phenol:chloroform purification. RNAs were resuspended in Ultrapure DNAse/Rnase Free Distilled Water (Thermo 10977035) RNA-seq: We used 0.5 μg of total RNA to purify polyadenylated mRNAs and to build an RNA library, using TruSeq Stranded mRNA Sample Prep Kit (Illumina, #RS-122-9004DOC), as recommended by the manufacturer. Directional libraries were checked for concentration and quality on DNA chips with the Bioanalyser (Agilent). More precise and accurate quantification were performed with sensitive fluorescent-based quantitation assays ("Quant-It" assays kit and QuBit fluorometer, Invitrogen). Samples were normalized to 2nM and multiplexed. After denaturation using 0.1N NaOH (5' at room temperature), the samples were diluted to 9pM and loaded on the flowcell. Sequencing was performed on the HiSeq 2500 sequencer (Illumina) in 65 bases V4 single-end mode
GEO Accession
GSM3350423