CUT&RUN Assay Kit #86652
- C&R
Product Information
Storage
实验步骤
Product Description
The CUT&RUN Assay Kit is designed to conveniently provide reagents needed to perform up to 8 (P size) or 24 (S size) digestion reactions from cells and is optimized for 5,000-250,000 cells per reaction. The kit has been optimized to work for all types of DNA binding proteins, including histones, transcription factors and cofactors. This kit has also been validated for fixed cells, primary cells, and tissues. A complete assay can be performed in as little as one day.
This kit provides an optional use of Yeast Normalization Spike-in DNA. It can be added to the CUT&RUN Stop Buffer to normalize signals across samples, correcting for technical variations among samples during the DNA purification, library preparation, and NGS steps. Also included is a Yeast Normalization Primer Set to facilitate sample normalization in downstream qPCR analysis.
In addition to the yeast spike-in normalization reagents described above, the kit provides additional key experimental controls to measure CUT&RUN assay success. These include the positive control Tri-Methyl-Histone H3 (Lys4) (C42D8) Rabbit Monoclonal Antibody #9751 and a negative control Rabbit (DA1E) Monoclonal Antibody IgG Isotype Control (CUT&RUN) #66362, both of which can be used for qPCR or Next Generation sequencing (NG-seq) analysis. PCR primer sets are provided for the human (#7014) and mouse (#7015) RPL30 gene locus to be used in conjunction with the control antibodies.
Specificity / Sensitivity
Background
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 cells or tissue to purified DNA, and has been shown to work with as few as 5,000-10,000 cells per assay (Figures 1-4). 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 of the sequencing depth that is required for ChIP-seq assays (1,2). Finally, the inclusion of yeast spike-in control DNA allows for accurate quantification and normalization, correcting for technical variations among samples during DNA purification, library preparation, and NGS steps.
Alternate Names
CnR; Cut & Run; CUT & Tag; cut and run; Cut and Run; cut and tag; CUT&RUN; CUT&Tag; cutandrun; cutandtag; histone modification
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