GSM9789686: 4T1, WT, rep4; Mus musculus; RNA-Seq
Total RNA was extracted using RNeasy Plus Mini Kit (Qiagen, 74134). RNA integrity was checked with Agilent 4200 TapeStation System. Following DNA contaminant removal and rRNA depletion, RNA-seq libraries were constructed using NEBNext Ultra II RNA Library Preparation Kit for Illumina. In brief, RNAs were fragmented at 94°C for 15 min. First and second strand cDNA synthesis was done using primers for random priming sites and unique molecular identifiers, which were incorporated into final cDNA. After adapter ligation, cDNAs were purified and enriched by PCR to generate sequencing libraries.