CUT&RUN 4X Stop Buffer #48105
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- C&R
Product Information
Product Usage Information
For the CUT&RUN assay, we recommend adding 37.5 μl CUT&RUN 4X Stop Buffer, 3.75 μl Digitonin Solution #16359, and 0.75 μl RNAse A (10 mg/ml) #7013 to 108 μl nuclease-free water (150 μl per reaction) right before use. Then incubate with the sample at 37°C for 10 min. An appropriate amount of Sample Normalization Spike-In DNA can be added to CUT&RUN 4X Stop Buffer, if desired.
Storage
Store CUT&RUN 4X Stop Buffer at -20°C. This product is stable for at least 36 months.
Product Description
The CUT&RUN 4X Stop Buffer provides enough reagent to support 24 CUT&RUN assays. This product is formulated for optimal performance in the CUT&RUN assay and each lot is tested and validated using the CUT&RUN Assay Kit #86652. This product should be diluted to 1X using nuclease-free water and an appropriate amount of Digitonin Solution #16359 and RNAse A (10 mg/ml) #7013 should be added right before use. Addition of Spike-In DNA is optional.
Background
Like the chromatin immunoprecipitation (ChIP) assay, Cleavage Under Targets and Release Using Nuclease (CUT&RUN) is a powerful and versatile technique used for probing protein-DNA interactions within the natural chromatin context of the cell (1-4). CUT&RUN provides a rapid, robust, and true low cell number assay for detection of protein-DNA interactions in the cell. Unlike the ChIP assay, CUT&RUN is free from formaldehyde cross-linking, chromatin fragmentation, and immunoprecipitation, making it a much faster and more efficient method for enriching protein-DNA interactions and identifying target genes. CUT&RUN can be performed in less than one day, from live cells to purified DNA, and has been shown to work with as few as 500-1,000 cells per assay (1,2). Instead of fragmenting all of the cellular chromatin as done in ChIP, CUT&RUN utilizes an antibody-targeted digestion of chromatin, resulting in much lower background signal than seen in the ChIP assay. As a result, CUT&RUN requires only 1/10th the sequencing depth that is required for ChIP-seq assays (1,2). Finally, the inclusion of simple spike-in control DNA allows for accurate quantification and normalization of target-protein binding that is not possible with the ChIP method. This provides for effective normalization of signal between samples and between experiments.
Alternate Names
C&R; CnR; CUT & RUN; cut and run; CUT and RUN; CUT&RUN; cutandrun; CUTandRUN; histone modification
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