GSM6203122: siRNA_1_ATAC; Homo sapiens; ATAC-seq
GSM6203122
In the assay, intact nuclei are treated with a hyperactive Tn5 transposase mutant which is able to simultaneously tag the target DNA with sequencing adapters to fragment the DNA in a process termed “tagmentation”. The ATAC-Seq Kit (Catalog No. 53150 Active Motif North America) was used for tagmentation and library preparation as per kit procedures. Briefly, cells were counted and aliquots of 50,000 to 100,000 cells were centrifuged at 500 x g for 5 minute at 4°C. Then the pellet was resuspended in 100 μl ice-cold ATAC Lysis Buffer. 50 μl of Tagmentation Master Mix was added to each sample and incubated at 37°C for 30 minutes in a thermomixer set at 800 rpm. This was followed by extraction and amplification of the tagmented DNA with library preparation using unique combinations of i7/i5 primers which are based on Illumina's Nextera adapters. Finally, the library was purified using SPRI bead clean-up.