title: GSM5291104: Microdissected Rat Renal cTALs, Sham Control_4_30mins, RNA-Seq, Transcriptome 01; Rattus norvegicus; RNA-Seq
library_construction_protocol: For bulk kidney, obstructed kidneys from animals were rapidly removed and be processed individually by homogenizing in Trizol, then transferred to a 2.0 mL microcentrifuge tube for RNA extraction and library construction (TruSeq Stranded mRNA Library Prep Kit, Illumina) according to the manufacturer's protocol. For microdissecting cortical collecting duct and cortical TAL,  total RNA were extracted using the Direct-zol™ RNA Kits (Zymo Research) and cDNA was synthesized by SMARTer V4 Ultra Low RNA kit (Clontech). The cDNA was then amplified for 15 PCR cycles, purified by AmPure XP magnetic beads (Beckman Coulter Genomics), and eluted with 15 μL Elution Buffer. The purified cDNA was further quantified by Qubit. Library size distribution was determined using the Agilent 2100 bioanalyzer using High-Sensitive DNA kit. The Nextera XT DNA Library Preparation Kits (Illumina) was used to generate the libraries with input of 1 ng. RNA libraries were prepared for sequencing using standard Illumina protocols