GSM8539204: Day 28 [scATAC-seq]; Mus musculus; ATAC-seq
Nuclei were isolated from flash-frozen mouse skin using an adaptation of 10x's nuclei isolation (CG000365) Library was performed according to the manufacter's instructions (single cell ATAC v2 protocol, 10x Genomics). Briefly, nuclei suspensions are incubated in a Transposition Mix.Barcoded Gel Beads, transposed nuclei, and master mix are loaded together with partitioning oil into the chip to generate the gel bead-in-emulsion (GEM). Once gel beads are dissolved, oligonucleiotides containing 1) P5 sequence, 2) a 16 nucleotide 10x barcode, and 3) a read 1N sequence are released and form barcoded single-stranded DNA. The pooled barcoded DNA 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, paired-end constructs with P5 and P7 sequences and a sample index were added.