title: GSM5916115: Control_EC_D3_rHDL_rep3_S28; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was isolated from FACS sorted ECs and Macrophagess from ischemic muscles using Arcturus PicoPure RNA isolation Kit (Applied Biosystems) Lexogen QuantSeq 3'mRNA-Seq Library Prep Kit-FWD (Lexogen, Vienna, Austria) was used to prepare 3′ RNA-Seq libraries. The double stranded DNA concentrations were quantified using a 5200 Fragment Analyzer (Part # M5310AA, Agilent Technologies), which gave similar concentrations for each sample that ranged from 1.7 to 4.3 ng/ μL. The molar concentration of cDNA molecules in the individual libraries was calculated from the DNA concentration and the region average size. Aliquots containing an equal number of nmoles of molecules from each library were pooled to give a pooled library with a concentration of 10 nM molecules. After library preparation, libraries were pooled and sequenced using single-end sequencing with 75 bp reads on an Illumina NextSeq500 instrument (Illumina, San Diego, CA).