GSM6199411: scRNASeq (10X), 6 populations from a pool of 33 C57BL/6 yolk sacs; Mus musculus; RNA-Seq
GSM6199411
Freshly collected yolk sacs were dissociated and separated into 6 pools that were hashtagged using MultiSeq reagents. 6 populations were sorted for those 6 pools, each with a unique hashtag. Populations were combined to run in a single 10X experiment. Library was performed according to the manufactorer's instructions (single cell 3' v2 protocol, 10x Genomics). Briefly, GCs were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI) and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR and the apropiated sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, paired-end constructs with P5 and P7 sequences and a sample index were added.