ATAC-seq experiments:
- After cell lysis and nuclei isolation, the nuclei pellets can be incubated with the following mix for 1 reaction:
| Tagmentation Buffer (2x) | 25 µl |
| Tagmentase loaded | 2.5 µl |
| Digitonin 1% | 0.5 µl |
| Tween20 10% | 0.5 µl |
| PBS | 16.5 µl |
| Nuclease-free water | 5 µl |
| Nuclei pellet* |
* The number of nuclei per reaction will depend on the ATAC-seq experimental design. Successful tagmentation with the proposed protocol has been performed on 50,000 nuclei per reaction.
- The reaction is then incubated 30 minutes at 37°C.
- The tagmentation reaction can then be stopped by addition of 250 µl of DNA Binding buffer from Diagenode MicroChIP DiaPure Columns (Cat. No. C03040001).
- The tagmented libraries can then be purified using the MicroChIP DiaPure Columns (Cat. No. C03040001), and amplified.
