Illumina HiSeq 4000 paired end sequencing; Hashing (sample multiplexing) of MCF7, PC3, DU145 and MDA-MB-231 cells and nuclei using different hashing methods
Hashing (sample multiplexing) of MCF7, PC3, DU145 and MDA-MB-231 cells and nuclei using different hashing methods
cells were trypsinized (0.05% trypsin-EDTA, Gibco, # 25300054) and pelleted at 200 x g for 5min. The cells were resuspended in 2 ml of culture medium, gently mixed with 8 ml of 1X PBS and put on ice. Afterwards, the cells were pelleted at 200 x g for 5min on 4°C and resuspended in 1 ml 1X PBS with 0.04% BSA.  MCF7, PC3, DU145 and MDA-MB-231 cells were maintained according to standard procedures in RPMI-1640 (Gibco, #21875034), F-12K (LGCstandards, ATCC 30-2004), EMEM (LGCstandards, ATCC 30-2003) and DMEM: F-12 (LGCstandards, ATCC 30-2006) medium respectively, supplemented with 10% fetal bovine serum (Gibco, #10082147) and 1% penicillin/streptomycin (100 U/ml and 100 μg/ml, respectively, Gibco, #15140122) at 37 °C with 5% CO2. RPMI-1640 medium used to culture the MCF7 cells was additionally supplemented with 0.01 mg/ml human recombinant insulin (Sigma Aldrich, #I3536). DNA was isolated according to 10x v3 user guides DNA library preparation was performed according to 10x v3 user guides