ATAC-Seq was performed as previously described22 (link) on 5,000–40,000 FACS-purified cells from 2–9 mice. In brief, cells were lysed in lysis buffer for 1 minute and transposed with Tagment DNA Enzyme 1 (Illumina) for 30 minutes. DNA was cleaned up using a MinElute DNA purification Kit (Qiagen), followed by barcoding and library preparation by the Nextera DNA Library preparation kit (Illumina) according to manufacturer’s guidelines and sequenced on an Illumina NextSeq500.
Protocol detail
ATAC-Seq on FACS-Purified Cell Samples
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Corresponding Organization : Rockefeller University
Other organizations : Keio University
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Variable analysis
independent variables
- Number of FACS-purified cells (5,000–40,000)
- Number of mice (2–9)
- Accessible chromatin regions identified by ATAC-Seq
- Lysis buffer composition
- Tagment DNA Enzyme 1 (Illumina) concentration and incubation time
- MinElute DNA purification Kit (Qiagen) for DNA cleanup
- Nextera DNA Library preparation kit (Illumina) for barcoding and library preparation
- Illumina NextSeq500 for sequencing
- Positive control: Not specified
- Negative control: Not specified
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