Single cell RNA sequencing of colorectal cancer with diverse tissue dissociation protocols or fresh vs. frozen cell preparations.. Single cell RNA sequencing of colorectal cancer with diverse tissue dissociation protocols or fresh vs. frozen cell preparations.

To evaluate 3 different tissue dissociation protocols or fresh vs. frozen cell preparations, we performed single-cell RNA sequencing on cancer or distant normal tissue dissociates from 2 colorectal cancer patients. Total 18,409 cells from 10 sample preparations were analyzed (5 primary colorectal cancer and 5 matched normal mucosa). The results suggest highly consistent cellular proportions were recovered with different sample preparation methods. Overall design: Ten batches of single cell RNA sequencing data were obtained from 2 patients, with 5 different sample preparations for primary colorectal cancer or matched normal mucosa respectively. For one patient, 3 different tissue dissociation protocols were applied for cancer and matched normal mucosa. For another freshly prepared or frozen-thawed cell stocks were tested for cancer and matched normal mucosa. After sequencing, we applied filtering criteria using nUMI, nGene and percent of mitochondrial genes to eliminate low-quality cells and 18,409 cells passed the filtering criteria. ****raw data files have not been provided due to patient privacy issues and will be available in EGA as controlled access****