Do I need to optimize the Tn5 tagmentation conditions for different cell types when performing CUT&Tag?

Our optimized tagmentation conditions for the CUT&Tag assay work well across multiple cell lines, both adherent and suspension. We recommend using 2 μL of CUT&Tag pAG-Tn5 (Loaded) #79561 in each 50 μL of tagmentation reaction. Using a larger volume of pAG-Tn5 in each reaction doesn't increase the number of reads, peak number, or signal-to-noise ratio, according to in our in-house tests. In fact, pAG-Tn5 activity is more critical to the success of the CUT&Tag assay than the amount of pAG-Tn5 used.

Please make sure to store the pAG-Tn5 enzyme properly at -20℃ and do not use expired pAG-Tn5, as activity does decline over time. The shelf life of CUT&Tag pAG-Tn5 (Loaded) #79561 is six months. Additionally, we have optimized the salt concentration in our tagmentation buffer to ensure efficient target-specific tagmentation and to limit non-specific tagmentation at open chromatin regions.

Last updated: April 24, 2024