CUT&Tag-IT™ Assay Kit
Rapid and robust genome-wide analysis of histone marks and transcription factor binding at lower sequencing depths
CUT&Tag-IT™ Assay Kit Overview
Cleavage Under Targets and Tagmentation (CUT&Tag) is a method to investigate genomic localization of histone modifications and some transcription factors that reveals interactions between proteins and DNA or identifies DNA binding sites for proteins of interest.
Unlike MNase-Seq or ATAC-Seq methods that target open chromatin and are therefore dependent on chromatin accessibility, CUT&Tag utilizes an antibody-based enzyme tethering strategy to target specific histone modifications or proteins to reveal chromatin-binding information that is specific to those sites or proteins of interest.
CUT&Tag is based on the same principles as ChIP-Seq, but with several changes to the protocol that are advantageous in certain situations. Instead of the sonication of fixed chromatin and immunoprecipitation steps performed in ChIP-Seq protocols, in CUT&Tag, fresh (not frozen) unfixed cells are bound to concanavalin A beads and the antibody incubation is performed with cells in their native state. Directly following antibody binding, the chromatin is digested and NGS libraries are prepared in a single step by tagmentation using the protein A-Tn5 (pA-Tn5) transposome enzyme that has been pre-loaded with sequencing adapters.
CUT&Tag can rapidly produce high-quality results from less starting material than ChIP-Seq, and enables robust analysis from lower sequencing depths, saving both time and money.
Learn more about CUT&Tag and how it worksCUT&Tag-IT™ Assay Kit Advantages:
- Compatible with as few as 5,000 cells
- Complete kit with optimized protocol
- Developed for histone marks and some transcription factors
- Sequencing-ready libraries without the laborious and costly steps of ChIP-Seq
- Low background signal enables lower sequencing depth
- No artifacts caused by formaldehyde crosslinking
How the CUT&Tag-IT™ Assay Works
CUT&Tag vs. CUT&RUN vs. ChIP-Seq
CUT&Tag-IT™ Assay Kit | CUT&RUN | ChIP-Seq | |
---|---|---|---|
Performed Under Native Conditions? | Yes | Yes | No |
Chromatin Fragmentation Method | Tn5-based tagmentation | MNase digestion | Sonication |
Cell Number Requirements | 5,000-500,000 cells | 500,000 cells | 1-10 million cells |
Sequencing Depth Required * | 2 million reads | 8 million reads | 20-50 million reads |
Integrated Library Preparation? | Yes, uses tagmentation | No, separate library prep required | No, separate library prep required |
Compatible Targets | Primarily histone modifications, some transcription factors and co-factors | Wide range of histone modifications, transcription factors, and co-factors | Wide range of histone modifications, transcription factors, and co-factors |
Workflow Length | 1-2 days | 1-2 days | 2-3 days |
* Kaya-Okur et al. Nature Communications (2019) 10:1930
CUT&Tag-IT™ Assay Kit Contents
The CUT&Tag-IT™ Assay Kit components are shipped at two temperatures, with one box on dry ice for components to be stored at -20°C, and a second box at room temperature for components that are not to be frozen and must be stored at 4°C. Please store components according to the storage conditions in the manual after first use. All reagents are guaranteed stable for 6 months from date of receipt when stored properly.
Reagents included:
- 5% Digitonin, store at -20°C
- Concanavalin A Beads, store at 4°C
- CUT&Tag-IT™ Assembled pA-Tn5 Transposomes, store at -20°C
- Tagmentation Buffer, store at -20°C
- 1X Binding Buffer, store at 4°C
- 1X Wash buffer, store at 4°C
- Dig-Wash Buffer, store at 4°C
- Antibody Buffer, store at 4°C
- Dig-300 Buffer, store at 4°C
- Guinea Pig Anti-Rabbit Antibody, store at -20°C
- Protease Inhibitor Cocktail, store at -20°C
- 0.5M EDTA, store at RT
- 10% SDS, store at RT
- 10 µg/µL Proteinase K, store at -20°C
- DNA Purification Columns, store at RT
- DNA Purification Binding Buffer, store at RT
- DNA Purification Wash Buffer, store at RT
- DNA Purification Elution Buffer, store at RT
- 3M Sodium Acetate, store at RT
- 10 mM DNTPs, store at -20°C
- 5X Q5 Buffer, store at -20°C
- Q5 High Fidelity DNA Polymerase (2U/µL), store at -20°C
- i5 Indexed Primer 1, store at -20°C
- i5 Indexed Primer 2, store at -20°C
- i5 Indexed Primer 3, store at -20°C
- i5 Indexed Primer 4, store at -20°C
- i7 Indexed Primer 1, store at -20°C
- i7 Indexed Primer 2, store at -20°C
- i7 Indexed Primer 3, store at -20°C
- i7 Indexed Primer 4, store at -20°C
- SPRI Beads, store at 4°C
Don’t forget to use an antibody validated for CUT&Tag!
Active Motif specializes in manufacturing high-quality antibodies to histones, histone modifications, chromatin proteins, and other factors, including a growing list of antibodies that we have experimentally validated in-house to work well in CUT&Tag assays. Check out our list of CUT&Tag-validated antibodies.
For a limited time, you can get a free antibody of your choice with every purchase of the CUT&Tag-IT™ Assay Kit. Learn more.
CUT&Tag-IT™ Assay Kit Data
Figure 1: CUT&Tag-IT Assay Kit data correlate well with ENCODE ChIP-Seq data.
Chromatin landscapes from human K562 cells are shown across a 868 kb segment of the human genome region (Chr16: 10,400-11,250) for H3K27me3 (dark blue), phospho-Pol2Ser2 (red), CTCF (orange), H3K4me3 (green), H3K9ac (light blue) H3K9me3 (purple), and H3K27ac (black) showing high concordance of CUT&Tag-IT Assay Kit signals from 100,000 cells compared with ENCODE ChIP-Seq data from at least one million cells. Antibodies used in the CUT&Tag assays for the following targets were: H3K27me3: cat. no 39155, CTCF: cat. no. 61311, H3K4me3: cat. no. 39159, H3K9ac: cat. no. 39917, H3K9me3: cat. no. 39161, H3K27ac: cat. no. 39133.
CUT&Tag-IT™ Assay Kit Documents
Order CUT&Tag-IT™ Assay Kit
We recommend using a CUT&Tag-validated antibody for best results. For a limited time, you can get a free antibody of your choice with every purchase of the CUT&Tag-IT™ Assay Kit. Learn more.
You might also be interested in:
- Comprehensive Guide to Understanding and Using CUT&Tag Assays
- ATAC-Seq Kit
- View Epigenetic Services
- Browse All Epigenetics Resources
US Pat. No. 10,689,643, EP Pat. No. 2999784 and related patents and applications
Name | Format | Cat No. | Price | |
---|---|---|---|---|
CUT&Tag-IT™ Assay Kit | 16 rxns | 53160 | ¥14,300 | Buy |
pA-Tn5 Transposase | 10 µg | 53161 | Get Quote |