GSM7050383: Male, WT, rep1, ATAC library [B1_100_ATAC]; Mus musculus; ATAC-seq
Flash frozen kidney pieces were thawed on ice and minced into small pieces (<1mm) with a sterile razor blade and then dounced in Nuclei EZ Lysis Buffer (Sigma). The tissue was dounced 15X loose, filtered with a 200uM filter, and then 5X tight, incubated for 5 mins on ice, filtered through 40uM filter and spun down at 500G x 5min at 4°C in a swinging bucket centrifuge. Supernatant was removed and nuclei pellet resuspended in Nuclei EZ lysis buffer, incubated an additional 5 mins on ice and spin down again. The final pellet was resuspended in Nuclei Buffer provided in 10X Chromium Single Cell ATAC Library Kit v1.1 and passed through a pre-wetted 5uM filter. The targeted number of nuclei were ascertained by Trypan Blue hemocytometer count then transposed and loaded onto a GEM H Chip as per the manufacturer's specifications. v1.1 (10x – PN:1000175) reagents and index plate were used to generate the snATACSeq libraries. Libraries were processed using 10X Genomics Manual CG000209 using 9 cycles for Sample Index PCR. Libraries were checked using Tapestation D5000 tapes before sending to BGI for PE 150 sequencing (DNBSEQ-G400). snRNA-seq + snATAC-seq; 10X Chromium Single Cell Multiome + Gene Expression protocol (CG000338 Rev E)