GSM5898054: KB; Homo sapiens; miRNA-Seq
Three cell lines, H357, KB, and Hep-2 were procured and grown in DMEM-F12, DMEM, and MEM growth media respectively, supplemented with 10% FBS and 1% antibiotics Pen-Strep under optimal conditions. Total RNA was extracted from the cell lines using the trizol method, yielding 37280ng (from H357), 54800ng (from KB), 1620ng (from HOK), and more than 5000ng (in Hep-2). The purity of isolated RNA was determined using a NanoDrop spectrophotometer and absorbance measurements at 260 and 280 nm. Total RNA of HOK were procured directly from ScienCell Research Laboratories, USA. An aliquot of each sample isolated from these cell lines were also tested on an Agilent RNA Bioanalyzer chip for RNA integrity number (RIN), which was found to be 10 for all cell lines, indicating intactness of the isolated RNA. The RNA libraries were constructed for sequencing following NEB Next Small RNA Sample Preparation protocol. After library preparation, they were sequenced on Illumina HiSeq 2500 with a 1x50 bp read size to obtain 60-85 million reads. Following the sequencing, the Illumina small RNA-Seq data was processed to create FASTQ files.