GSM4475044: standard diet 2m-3; Mus musculus; RNA-Seq
The total RNA was extracted from whole kidneys using the Nucleospin® RNA Isolation Kit (MACHEREY-NAGEL, Germany). The libraries were prepared for 100bp paired-end sequencing using TruSeq RNA Sample Prep Kit (Illumina, CA, USA). Total 1μg of RNA molecules were purified and fragmented and synthesized as single-stranded cDNAs through random hexamer priming. By applying this as a template for second strand synthesis, double-stranded cDNA was prepared. After sequential process of end repair, A-tailing and adapter ligation, cDNA libraries were amplified with PCR (Polymerase Chain Reaction). Quality of these cDNA libraries was evaluated with the Agilent 2100 BioAnalyzer (Agilent, CA, USA). They were quantified with the KAPA library quantification kit (Kapa Biosystems, MA, USA) according to the manufacturer's library quantification protocol. Following cluster amplification of denatured templates, sequencing was progressed as paired-end (2×100bp) using Illumina HiSeq2500 (Illumina, CA, USA).