|
Single cells were obtained as gel bead-in-emulsions (GEMs) using the 10X Genomics (Pleasanton, CA, USA) Chromium platform based on the manufacturer's protocol. Briefly, cell suspensions of skin tissue with reverse transcription master mix were loaded onto the 10X Genomics Single Cell 3' chip and partitioned into GEMs with gel beads precoated with oligonucleotides containing a poly-dT primer sequence to capture released mRNAs, a 16-bp 10X barcode for cell indexing, and a 10-bp unique molecular identifier (UMI) to distinguish between transcripts. After reverse transcription, barcoded cDNAs for each sample were amplified to generate a sufficient volume for library construction using the Single Cell 3' Reagent Kit (v3 chemistry). Libraries were subjected to 150-bp paired-end sequencing performed by Novogene (Beijing, China) on a NovaSeq 6000 system (Illumina, San Diego, CA, USA). |
ssRNA-Seq |
TRANSCRIPTOMIC SINGLE CELL |
unspecified |
PAIRED
|
