Illumina NovaSeq 6000 sequencing; Adult mouse SVZ niche at the single cell resolution
Adult mouse SVZ niche at the single cell resolution
Cervical dislocation and microdissection of lateral walls of the SVZ. Papain digestion complemented with mechanical dissociation followed by filtration, myelin removal with a Percoll gradient, and red blood cell lysis. Cells capture, lysis & RNA extraction were performed using the 10x V3 Chromium technology as recommended by the manufacturer. Around 12,000 cells by replicate were loaded in each lane of the Chromium 3′chip.  Barcoding and library preparation were performed with the 10x Chromium system using version 3 chemistry (Dual index SI_TT_B1 for SVZ CTRL and SI-TT-B2 for SVZ_4Gy) ), following the manufacturer's protocol. cDNA was synthesized and amplified as per manufacturer's protocol and amplified to construct an Illumina sequencing library. cDNA and libraries were checked for quality on using Agilent with DNA HS chip.