For the CUT&Tag assays, we recommend adding 120 μL 10X High Salt Wash Buffer (CUT&Tag), 12 μL 100X Spermidine #27287, 6 μL Protease Inhibitor Cocktail (200X) #7012, and 30 μL Digitonin Solution #16359 to 1,032 μL of nuclease-free water (1.2 mL per reaction) right before use. Please keep on ice during use.
Storage
Store 10X High Salt Wash Buffer (CUT&Tag) at 4°C. This product is stable for at least 12 months.
Product Description
The 10X High Salt Wash Buffer (CUT&Tag) provides enough reagent to support 24 CUT&Tag assays. This product is formulated for optimal performance in the CUT&Tag assays and each lot is tested and validated using the CUT&Tag Assay Kit #77552.
Background
Similar to Cleavage Under Targets and Release Using Nuclease (CUT&RUN), Cleavage Under Targets and Tagmentation (CUT&Tag) is a powerful technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1-3). CUT&Tag has many of the same advantages as the CUT&RUN assay in that it provides a rapid, robust, and true low cell number protocol for detection of protein-DNA interactions in the cell. In addition, the CUT&Tag assay adds an in situ adaptor DNA ligation step carried out by the pAG-Tn5 enzyme, in which an adaptor DNA is ligated directly to antibody-targeted chromatin DNA fragments in the cell. As a result, subsequent DNA library preparation is much faster and easier than library preparation following the CUT&RUN assay, free from DNA end repair, A-tailing, and adaptor ligation in vitro. CUT&Tag works very well for analyzing histone modifications, in addition to mapping some transcription factor and cofactor binding.
