GSM5725854: MCF-12A control replicate 2; Homo sapiens; RNA-Seq
RNA was isolated with Qiagen RNeasy plus mini Kit. Total RNA was first evaluated for its quantity, and quality, using Agilent Bioanalyzer 2100. A RIN (RNA Integrity Number) of eight or higher was required to pass the quality control. One hundred to 250 nanograms of total RNA per sample were used for library preparation using KAPA RNA HyperPrep Kit (Roche) for Illumina NGS platform. Each indexed library was quantified and its quality accessed by Qubit and Agilent Bioanalyzer, and 6 libraries were pooled in equal molarity. 5µL of 4nM pooled libraries were denatured, neutralized and a final concentration of 1.5 pM of pooled libraries was loaded to Illumina NovatSeq 6000 for 75bp single-read sequencing.