GSM8901878: CLE_Patient1_non-lesional; Homo sapiens; RNA-Seq
For single-cell RNA-seq: 4 or 5 mm punch biopsies were bisected into four pieces and digested with Dispase II (10 mg/mL) in RPMI supplemented with 10% FBS, 1% L-Glutamine, and 1% Penicillin/Streptomycin for 45 minutes. The tissue was then cut into finer pieces with a razor and placed in 0.5 mL/mL Liberase TH with 40 ug/mL DNase I for another 45 minutes. Tissue and media were passed through a 70 uM strainer, counted, and suspended in 0.04% BSA and PBS. For spatial transcriptomics: FFPE tissues were sectioned onto glass slides. Deparaffinization, H&E staining, imaging, and decrosslinking were conducted following the 10x Genomics protocol CG000520. The Visium assay was performed using CytAssist technology according to protocol CG000495. Library construction protocol: For single-cell RNA-seq: 10x Genomics 5'v2 kit was used for library preparation. Samples were sequenced at a minimum depth of 20,000 read pairs per cell. For spatial transcriptomics: Following the transfer of gene expression probes from tissue to the Visium slide using CytAssist, the probes were extended, amplified, and made into libraries for sequencing. Sequencing protocol: Single-cell libraries were sequenced on an Illumina platform to a minimum depth of 20,000 read pairs per cell. Spatial transcriptomics libraries were sequenced using paired-end 150 base reads on an Illumina NovaSeq instrument.