DT17043_L
single-cell RNA-seq of human tissue: pancreatic ductal adenocarcinoma: Sample DT17043
"The tissue specimen was collected, minced, and enzymatically digested in 10% FBS/DMEM supplemented with 2.5 mg/mL Collagenase D, 0.5 mg/mL Liberase DL, and 0.2 mg/mL DNase I for 45 minutes at 37oC with agitation. The digest was quenched by adding PBS containing 2% FBS, filtered through a 100 uM cell filter, and centrifuged to collect single cells. Fluorescence activated cell sorting was used to isolate viable (DAPI-negative) cells. Sorted cells were collected in PBS containing 20% FBS. Sorted cells were washed and resuspended in PBS containing 0.04% BSA. Up to 12,000 cells were loaded per lane on 10X Chromium microfluidic chips for single-cell capture. Single-cell barcoding and library preparation were performed using the 10X ChromiumTM version 2 chemistry, according to the manufacturer's protocol. The library was sequenced on an Illumina HiSeq 4000 instrument, and scRNA-seq reads were processed into FASTQ files Cell Ranger pipeline (v1.3, 10X Genomics). *R1_001.fastq.gz files contain the cell barcode (first 16 nt) and UMI barcode (next 10 nt). *R2_001.fastq.gz files contain the 98 bp RNA sequence. *I1_001.fastq.gz files contain the 8 bp index read."
DT17043_L
CSHL-phs001840
DT17043_L