title: GSM4698711: MC B7low PBS 1; Mus musculus; RNA-Seq
library_construction_protocol: Flow cytometrically identified MCs were sorted into 1.5 mL Eppendorf tubes containing 5 uL of 1xTCL buffer (Qiagen). After sorting, tubes were immediately sealed, subjected to a quick spin on a benchtop microcentriguge, and frozen at -80 C. SmartSeq2 libraries were prepared according to the SmartSeq2 protocol (Picelli et al, 2013, 2014) with some modifications (Trombetta et al, 2014). Briefly, total RNA was purified using RNA-SPRI beads. Poly(A)+ mRNA was converted to cDNA which was then amplified. cDNA was subject to transposon-based fragmentation that used dual-indexing to barcode each fragment of each converted transcript with a combination of barcodes specific to each sample. Barcoded cDNA fragments were then pooled prior to sequencing. Sequencing was carried out using an Illumina NextSeq500 as paired- end 2x25bp with an additional 8 cycles for each index.