GSM4766628: Sample 1_CD34+ cells Delta1ext-IgG normoxia bulkRNA rep1; Homo sapiens; RNA-Seq
Total RNA was extracted using the RNeasy Plus Mini Kit (Qiagen) according to the manufacturer's instructions. The genomic DNA elimination columns contained in the kit were used to eliminate possible DNA contamination during the extraction. The sequencing libraries were constructed from 10 pg–10 ng of total RNA using the Takara Bio USA's SMART-Seq V4 Ultra Low Input RNA Kit for Sequencing (Cat # 634888) following the manufacturer's instructions. Full-length cDNA synthesis was performed using oligo-dT and SMART oligos, and the Illumina sequencing adaptors and barcodes incorporated into the final library by PCR. The fragment size of the RNAseq libraries was verified using the Agilent 2100 Bioanalyzer (Agilent) and the concentrations determined using the Qubit instrument (Thermo Fisher Scientific). The libraries were loaded onto the Illumina HiSeq 3000 machine (Illumina) for paired end 75 bp read sequencing and generated about 50 M reads per sample. The.fastq files were generated using the bcl2fastq software (Illumina) for further analysis.