GSM2840135: WT_D4MI_CD45P; Mus musculus; RNA-Seq
Hearts were harvested, flushed, and enzymatically digested to release single cells. Cells were labeled with DAPI and CD45 antibody and all live CD45+ cells were collected by flow cytometry. Cells were encapsulated using the inDrop platform, into droplets on ice and lysed in 4nL microfluidic droplets using a final concentration of 0.4% NP-40. Single cell lysates were subject to reverse transcription at 50C without purification of RNA as previously described (Klein et al., Cell 2015). Cells were barcoded using the inDrop platform (Klein et al., Cell 2015), which makes use of the CEL-Seq protocol for library construction (Hashimshony et al., Cell Reports 2012). Microfluidic inDrop single cell libraries were prepared as described in Zilionis et al. Nature Protocols 2017 (doi:10.1038/nprot.2016.154). Sequencing was performed on an Illumina HiSeq 2500.
GEO Accession
GSM2840135