title: GSM8284866: Monocy_4T1_In_vivo_Cas9; Mus musculus; RNA-Seq
library_construction_protocol: FACS-purified cells were lysed immediately upon direct sorting in 150 µL of extraction buffer from the Arcturus PicoPure RNA isolation kit (Thermo Scientific). The quality of the RNA isolated through the PicoPure kit was determined with a TapeStation RNA High sensitivity Screen Tape (Agilent). Due to the low concentration of total RNA the SMARTer® Stranded Total RNA-Seq Kit v3 -Pico Input Mammalian (Takara Bio) was used to prepare sequencing libraries. Briefly, total RNA samples (0.27–8 ng) were mechanically fragmented (4 minutes) and reverse-transcribed using random priming into double-stranded cDNA in the presence of a template switch oligo (TSO). When the reverse transcriptase reaches the 5' end of the RNA fragment, the enzyme's terminal transferase activity adds non-templated nucleotides to the 3'