GSM4891841: 6 hrs after IRI; Mus musculus; RNA-Seq
Kidneys were dissociated with liberase TM, hyaluronidase, DNaseI at 37oC for 40 min, followed by incubation with 0.25% trypsin EDTA at 37oC for 30 min. Trypsin was inactivated using 10% fetal bovine serum in PBS. Cells were then resuspended in PBS supplemented with 0.01% bovine serum albumin then filtered through 40 µm strainer and utelized for Drop-seq DNA polymerase for pre-amplification step (1 cycle of 95°C for 3 min, 15-17 cycles of 98°C for 15 sec, 65°C for 30 sec, 68°C for 4 min and 1 cycle of 72°C for 10 min) was replaced by Terra PCR Direct Polymerase (#639271, Takara). The other processes were performed as described in the original Drop-seq protocol. Sequencing was performed using Hiseq X ten. single-cell RNA-seq (Drop-seq)