- 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|
|Nuclease-free water||5 µl|
* 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.