single-cell whole genome sequences of homo sapiens: IMR-90 primary cell
Single cells were isolated using the CellRaft AIR system (Cell Microsystems). The CellRaft array was prepared following the manufacturers instruction and washed three times for 3 minutes with warm PBS before addition of cell suspension. 1000 cells were seeded in 1ml for one CellRaft array. Cells were allowed to settle for 3-4 hours at 37C before isolation. Before isolation the raft was carefully washed with warm medium to remove potential debris and dead cells. Individual rafts containing one cell were deposited in PCR tubes with 2.5ul PBS and stored at -80C until further processing. Libraries for whole genome sequencing (WGS) were prepared according to the previously described scMDA protocol using a NEBNext Ultra II FS kit (NEB). For quality control purposes, a locus drop out test (LDO) was performed using primers and SYBR reagent [34]. Library size was assessed with a TapeStation (Agilent) using HS5000 screen tape for quality control of cDNA and HS1000 screen tape for all final libraries. All RNA-Seq and WGS libraries were sequenced on an Illumina HiSeq platform by Novogene Corp Inc., CA.