GSM4154965: black650_fatp2KO; Mus musculus; RNA-Seq
RNA was isolated using standard QIAzol protocol (Cat No./ID 79306). After RNA isolation, we cleaned up the RNA using the Qiagen RNeasy kit following the manufacturer's protocol (Cat No./ID 74104). Quality control checks of the RNA were performed to define purity and concentration. We used a nanodrop to assess the quantity and quality for each sample, ensuring the quantity was between 50-250ng/µl and the 260/280 was between 1.95-2.10. We ensured no RNA degradation occurred using Agarose Gel Electrophoresis for each sample. Lastly, before sequencing Novogene performed additional quality and quantity assessments of the RNA samples using an Agilent 2100 instrument and gel electrophoresis. RNA-Seq libraries were prepared using Illumina TruSeq® RNA sample preparation kit as per the manufacturer's protocol. Library construction, additional quality assessment, and sequencing were performed by Novogene Corporation, Ltd. Library concentration was quantified with a Qubit 2.0 fluorometer from Life Technologies and samples diluted to 1ng/µL before checking insert size using a bioanalyzer (Agilent 2100 Bioanalyzer) and quantifying to greater accuracy using qPCR. Using paired-end sequencing at a depth of 40 million reads or greater per sample was completed on final libraries. We used paired-end sequencing for RNA-sequencing experiments.