title: GSM5051350: aSMA periosteum 2 days and periosteum 13 weeks (mixed samples); Mus musculus; RNA-Seq
library_construction_protocol: Custom designed Drop-Seq barcodes from Integrated DNA Technologies (IDT) were delivered into each well of each 384 well plate. All primers in one well shared the same unique cell barcode and billions of different unique molecular identifiers (UMIs). An Echo 525 liquid handler was used to sequentially dispense lysis buffer, primers (custom designed Drop-Seq barcodes from Integrated DNA Technologies), and reaction reagents totaling 1 microliter into each well in the plate for the cell lysis and cDNA synthesis using a modified Drop-Seq protocol. Following cDNA synthesis, the contents of each well were collected and pooled into one tube using a Caliper SciClone Liquid Handler. After treatment with exonuclease to remove unextended primers, the cDNA was PCR amplified for 13 cycles and the cDNA was fragmented and amplified for sequencing using a Nextera XT DNA sample prep kit (Illumina) using custom primers (see below) that enabled the specific amplification of only the 3′ ends. scRNA-seq