title: GSM3097061: N_plus_CTX_rep1; Mus musculus; RNA-Seq
library_construction_protocol: Mice were euthanized and injected muscles (or uninjected controls) were carefully harvested. Muscle was minced in digestion media containing Collagenase II, DNase I, and dispase for 55 minutes and subsequently filtered over a 70μm filter. Cells were purified using a 37.5% Percoll gradient to generate single cell suspensions. Cells were stained and sorted on the basis of 7AAD-CD45+CD64+MerTK+ cells on an BD Aria I. Bulk sorted cells were prepared for single-cell capture using a 10X Genomics Chromium platform. cDNA was prepared on a BioRad T1000 overnight and double stranded DNA libraries were prepared. After a final Ampure XP (Beckman Coulter) bead cleanup, libraries were quantified using Qubit fluorescence and assessed using the Advanced Analytics Fragment Analyzer using standard reagents. Absolute concentration of libraries was achieved using the Kapa Biosystems Universal qPCR protocol according to the manufacturer. Following qPCR, libraries were diluted and denatured to 8 pM for loading onto a HiSeq2500 in RAPID mode with 26 x 98 paired end run parameters.
attributes.tag: GEO Accession
attributes.value: GSM3097061