Description
Product Usage Information
Please refer to CUT&Tag Assay Kit # 77552 for detailed use of this enzyme in the CUT&Tag assay. After cell permeabilization and primary and secondary antibody binding, resuspend cells in 50 μL of High Salt Digitonin Buffer containing 2 μL of pAG-Tn5 Enzyme (1:25 dilution). Incubate cell samples at room temperature for 1 hour, wash cells with High Salt Digitonin Buffer, and then perform the chromatin tagmentation.
Storage
Supplied in 22 mM HEPES pH 7.4, 44 mM NaCl, 44 μM EDTA, 0.4 mM DTT, 0.04% Triton X-100, and 50% glycerol. Store at -20°C and do not aliquot. This product is stable for 6 months.
Specificity / Sensitivity
Species Reactivity: All Species Expected
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 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 . CUT&Tag works very well for analyzing histone modifications, in addition to mapping some transcription factor and cofactor binding.
Alternate Names
C&T; CnT; CUT & Tag; cut and tag; CUT and Tag; CUT&Tag; cutandtag; histone modification
