NextSeq 500 paired end sequencing; GSM3109654: Sample 1_PWD12_WoundDermis_SingleCells_Fluidigm_Cell_1; Mus musculus; RNA-Seq

Sample 1_PWD12_WoundDermis_SingleCells_Fluidigm_Cell_1

We report transcriptomes of pre-sorted skin wound dermal cells. Post-wounding day (PWD) 12, 15 and 21 Zombie-neg;tdTomatoHi cells were  FACS sorted from Sm22-Cre;TdTomato mice. Overall design: Examination of single cell heteregeneity in large skin wounds on PWD 12, 15 and 21

Single-cell transcriptomics confirms heterogeneity of contractile cells in large skin wounds

Submitted on 06-DEC-2018

Skin wound tissues were harvested and dissegragated into single cell suspensions with collagenase. Cells were FACS sorted as viable tdTomatoHi. Pre-sorted, viable tdTomatohi single cells were re-suspended at appropriate concentrations in DMEM supplemented with 10% FBS, antibiotics and antifungals, diluted with suspension reagent for attribution of optimal buoyancy, and loaded onto a large 17-25μm 96-well microfluidic IFC (Fluidigm) for single cell capture in the automated C1 system for single-cell genomics (Fluidigm). Lysis, RT and cDNA pre-amplification were performed in loco (protocol 1.773x) with ultra-low input RNA reagents as suggested per manufacturer (Clonetech). RNA spike-in controls were omitted. cDNA concentrations were estimated using Qubit 2.0 (Thermo) and cDNAs with concentration ≥ 1.0 ng/μl were used for downstream library preparation. Libraries were amplified using the Nextera XT v2 Index Kit (Illumina). Quality control on multiplexed libraries was performed using the Agilent Bioanalyzer and quantification was performed using KAPA for Illumina Sequencing Platforms (Illumina). Multiplexed libraries were sequenced as paired-end on an Illumina Next-Seq500 platform (Illumina). SMART-seq2 - Fluidigm.

Single-cell transcriptomics confirms heterogeneity of contractile cells in large skin wounds

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