Single cell ATAC-seq of 10 ug/ml anti-IgM-stimulated mouse B cell: adult spleen
Single cell ATAC-seq was performed using splenic B cells in the presence or absence of anti-IgM (1 hour, 10 μg/ml). The single cell ATAC-Seq libraries were prepared using C1 system (Fluidigm) and Nextera DNA Library Preparation Kit (Illumina) following the previous research (Buenrostro et al., Nature, 2015) with a modification of second PCR condition as follows; 1 cycle of 72ºC for 5 minutes, and 98ºC for 30 seconds; 13 cycles of 98ºC for 10 seconds, 63ºC for 30 seconds, and 72ºC for 1 minute. 53 and 50 libraries of control and anti-IgM-stimulated B cells originated from single cell were pooled, respectively. To remove primer-dimer, the pooled libraries were purified using the same volume of AMPure XP beads three times. 101 bp pair end multiplex sequencing was performed by HiSeq2500 (Illumina).