Single Cell RNA-Seq of Ax3, Ax4, Ax3 SET1 KO and Ax4 SET1 KO
Cells were isolated in a SMARTer ICELL8 3 DE Chip using the ICELL8 cx Single Cell System. Wells of the 3 DE Chip contained pre-printed oligonucleotides possessing well specific barcodes and UMIs connected to a polydT region for hybridisation with polyadenylated transcripts. After identification of wells containing cells, cells were lysed and components for reverse transcription (RT) and cDNA amplification were dispensed into chosen wells. After RT-PCR, products from separate wells were collected into a single sample, concentrated and purified according to the manufacturers instructions. Samples were prepared for sequencing using a Nextera XT Library Prep Kit and sequenced over 4 lanes of an entire flow cell using a NextSeq High-Output kit (2 x 75bp reads). One read was used to sequence the well barcode and transcript UMI, with the second reading the 3 end of the transcript itself.