GSM4567350: M1WT_524; Mus musculus; RNA-Seq
Whole kidneys from the anatomical left were immediately excised from euthanized WT and CTRP1-deficient mice and flash frozen in liquid nitrogen. Total RNA was isolated using Trizol reagent (Thermo Fisher Scientific, Carlsbad, CA.)  per the manufacturer's protocol. RNA samples were subjected to DNASE I (New England Biolabs, Ipswwich, MA.) digestion to remove genomic DNA contamination using the manufacturer's protocol. A 3M (pH 5.2) sodium actetate (Quality Biological, Gaithersburg, MD.) precipation was performed after DNASE I digestion to purify RNA samples.  In brief, sodium acetate was added at 1/10 the volume of the sample followed by the addition of 2.5 volumes of ice cold 100% ethanol and mixed throroughly. The samples were incubated at -80 ° C overnight. The samples were centrifuged at 12,000 x g for 20 minutes at 4° C and the supernantant was decanted. The RNA pellets were washed twice using 1ml of ice cold 75% ethanol and centrifuged at 12,000 x g at 4° C. A quick spin was performed to removed any traces of residual ethanol in the pellet using a very fine pipette tip. The pellets were air dryed and reconsitituted in sterile filtered molecular biology grade water (Quality Biological, Gaithersburg, MD). The RNA quality number (RQN) of total RNA was determined for each sample before RNA-seq analysis was performed by Novogene. Quality-controlled total RNA was purified to mRNA with poly-T magnetic beads, fragmented, and then reverse transcribed into cDNA.  Fragments of 150~200bp were selected and the library PCR amplified.
GEO Accession
GSM4567350