GSM3351357: wt; Mus musculus; RNA-Seq
Sorted cells were counted and assessed for viability with Trypan Blue using a Countess II automated counter (Invitrogen), and adjusted at a concentration of 800-1000 cells/µL. Final cell viability estimates ranged between 80-93%. Single cell suspensions were loaded onto 10X Genomics Single Cell A Chips (10x Genomics, PN-120236) along with the reverse transcription (RT) master mix as per the manufacturer's protocol for the Chromium Single Cell 3' Reagent kits V2. (10X Genomics; PN-120237), to generate single cell gel beads in emulsion (GEMs) and sequencing libraries. Chromium Single Cell 3' Library & Gel Bead Kit v2 (10xGenomics) using manufacturer protocol The Single Cell 3' Protocol produces Illumina-ready sequencing libraries. A Single Cell 3' Library comprises standard Illumina paired-end constructs which begin and end with P5 and P7. The Single Cell 3' 16 bp 10x Barcode and 10 bp UMI are encoded in Read 1, while Read 2 is used to sequence the cDNA fragment. Sample index sequences are incorporated as the i7 index read. Read 1 and Read 2 are standard Illumina sequencing primer sites used in paired-end sequencing. Rapid mode as follows: 26 bp (Read 1); 8bp (i7 Sample Index); 0bp (i5 Index); 98bp (Reads 2).The BCL data will include the paired-end Read 1 (containing the 16 bp 10x Barcode and 10 bp UMI) and Read 2 and the sample index in the i7 index read.
GEO Accession
GSM3351357