GSM8843217: EmxDot E16.5 WT 469.1; Mus musculus; RNA-Seq
The dorsal telencephalon was extracted from the embryos and the cells were dissociated using the Worthington papain dissociation system and resuspended in PBS. The library was performed according to the manufacturer's instructions (single cell 3' v3.1 dual index protocol, 10x Genomics). Briefly, the dissociated brain cells resuspended in PBS were added to the master mix and loaded, together with gene expression beads and partitioning oil, into the chip and to generate the gel-bead emulsion using the Chromium machine. The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Illumina R1 primer sequence, a unique molecular identifier (UMI), and 10x barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the fragments with the appropriate sizeswere selected using SPRIselect beads for subsequent library construction. During the library construction Illumina R2 sequence, paired-end constructs with P5 and P7 sequence, and a sample (dual) index were added.