Illumina HiSeq 2500 sequencing; Bulk RNA-seq validation experiments for "High dimensional single cell analysis predicts response to anti-PD-1 immunotherapy"
Bulk RNA-seq validation experiments for "High dimensional single cell analysis predicts response to anti-PD-1 immunotherapy"
CD14+CD16-HLA-DRhiLin- (CD3, CD4, CD19, CD45RO) monocytes were sorted from frozen PBMC form blood samples from HD (healthy donor), R (responder) and NR (non-responder) at baseline. Representative samples (n=4, each) of responders/non responders/ and healthy donors were selected from archival samples stored in the dermatology biobank according to the same clinical criteria used in the discovery and validation cohorts for CyTOF and FACS analysis. RNA was isolated using the RNA plus micro kit (Qiagen). We used the SMARTer Stranded total rna-seq pico kit (Clontech) for RNA preparations. Libraries for sequencing were generated with the TruSeq RNA Sample Preparation kit (Illumina).
Experimental Factor: disease
metastatic melanoma
Experimental Factor: response to treatment
non-responder