GSM5592627: MC-SOX9-GFPhi_rep2; Mus musculus; RNA-Seq
Total RNA was isolated using QIAGEN RNeasy Mini kit and quantified using Qubit fluorometer (Thermo Fisher Scientific). Quality of the isolated RNA was checked using Agilent Bioanalyzer 2100. Universal Plus mRNA-Seq Library with NuQuant (TECAN) was used to generate stranded RNA-seq libraries. Briefly, poly(A) RNA was selected followed by RNA fragmentation. Double stranded cDNA was generated thereafter using a mixture of random and oilgo(dT) priming. The library was then constructed by end repairing the cDNA to generate blunt ends, ligation of Unique Dual Index (UDI) adaptors, strand selection and PCR amplification. Different adaptors were used for multiplexing samples in one lane. Quality of the library was checked using Agilent Bioanalyzer 2100. Sequencing was performed on Illumina Hiseq3000 with single end 50 base pair reads. Universal Plus mRNA-Seq Library with NuQuant (TECAN)