hotspot-10-25 KO R1
[1] cells are maintained in RPMI 1640 medium supplemented with FBS (10% v/v) and pen/strep (1% v/v, Sigma-Aldrich P4333). [2] lentivirus expressing pgRNA is used to infect the cells, and the cells are sorted at day 8 post infection. [3] cells are lysed as single cell in the droplet, and poly A+ RNA is captured by oligo-dT on the beads. [4] poly A+ RNA is reverse transcribed using Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV RT), then the cDNA is tagmented and amplified following Illumina Nextera DNA preparation kit. [5] drop-seq