

CUTANA™ pAG-Tn5 for CUT&Tag Workflows
{"AddThisServiceButtonMeta":"","add_this":[{"annotation":"","service":"facebook"},{"annotation":"","service":"email"},{"annotation":"","service":"print"},{"annotation":"","service":"twitter"},{"annotation":"","service":"linkedin"}],"add_to_wishlist_url":"/wishlist.php?action=add&product_id=764","availability":"","bulk_discount_rates":[],"can_purchase":true,"cart_url":"https://www.epicypher.com/cart.php","category":["Epigenetics Kits and Reagents","Epigenetics Kits and Reagents/CUTANA™ CUT&Tag Assays"],"custom_fields":[{"id":"1005","name":"Internal Comment","value":"extra boxes in bottom shelf of Venom"}],"customizations":[],"description":"<div class=\"product-general-info\">\n <ul class=\"product-general-info__list-left\">\n <li class=\"product-general-info__list-item\">\n <strong>Type: </strong>Transposase\n </li>\n <li class=\"product-general-info__list-item\">\n <strong>Mol Wgt: </strong>191 kDa\n </li>\n </ul>\n <ul class=\"product-general-info__list-right\">\n <li class=\"product-general-info__list-item\">\n <strong>Host: </strong><em>E. coli</em>\n </li>\n <li class=\"product-general-info__list-item\">\n <strong>Epitope Tag: </strong>None\n </li>\n </ul>\n</div>\n<div class=\"service_accordion product-droppdown\">\n <div class=\"container\">\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel current\">\n <h3 class=\"sub-title1\">Description</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description-specific\"\n style=\"display: block\"\n >\n <p>\n Available in 50 and 250 Reaction pack sizes. CUTANA pAG-Tn5 is the\n key reagent for performing efficient mapping of chromatin features\n in Cleavage Under Targets and Tagmentation (CUT&Tag) [1]. CUT&Tag\n offers significant improvements in signal to noise at reduced cell\n inputs and sequencing depth compared to ChIP-seq. As a fusion of\n Proteins A and G to a highly active\n <em>E. coli</em> transposase mutant (Tn5), CUTANA pAG-Tn5 is\n compatible with target antibodies from a broad spectrum of host\n species. It also comes without an epitope tag, making it suitable\n for tag-mediated CUT&Tag (e.g. FLAG, HA, TY1, V5, etc.). This\n product is highly purified to remove contaminating\n <em>E. coli</em> DNA, which can be tagmented during storage and\n complicate analysis at low cell numbers. For superior normalization\n as well as antibody validation and reaction monitoring,\n <a\n href=\"https://www.epicypher.com/products/nucleosomes/snap-cutana-spike-in-controls\"\n >SNAP-CUTANA™ nucleosome spike-ins</a\n >\n are recommended. The Tn5 comes charged with mosaic adapters and is\n ready to be used in CUT&Tag.\n </p>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel current\">\n <h3 class=\"sub-title1\">Validation Data</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description-specific\"\n style=\"display: block\"\n >\n <p>\n CUT&Tag was performed on 100k K562 cells with 0.5 µg of either IgG\n (EpiCypher\n <a\n href=\"/products/nucleosomes/snap-cutana-spike-in-controls/cutana-rabbit-igg-cut-run-negative-control-antibody\"\n >13-0042</a\n >), H3K4me3 (EpiCypher\n <a\n href=\"/products/antibodies/snap-chip-certified-antibodies/histone-h3k4me3-antibody-snap-chip-certified-cutana-cut-run-compatible\"\n >13-0041</a\n >), or H3K27me3 (<a\n href=\"https://www.thermofisher.com/antibody/product/H3K27me3-Antibody-clone-G-299-10-Monoclonal/MA5-11198\"\n >ThermoFisher MA5-11198</a\n >) antibodies using CUTANA™ pAG-Tn5 (1:20 dilution) following the\n EpiCypher Direct-to-PCR CUT&Tag <a href=\"/protocols\">protocol</a>.\n Libraries were run on an Illumina NextSeq2000 with paired-end\n sequencing (2x50 bp). Sample sequencing depth was 0.3 million reads\n (IgG), 1.7 million reads (H3K4me3), and 3.8 million reads\n (H3K27me3). Data were aligned to the hg19 genome using Bowtie2. Data\n were filtered to remove duplicates, multi-aligned reads, and\n blacklist regions.\n </p>\n\n <section class=\"image-picker\">\n <div class=\"image-picker__left\">\n <div\n class=\"image-picker__main-content_active image-picker__main-content\"\n >\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/proteins/15-1017-protein-gel-data.jpeg\"\n target=\"blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"15-1017-protein-gel-data\"\n src=\"/content/images/products/proteins/15-1017-protein-gel-data.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n ></a\n >\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\"\n ><strong>Figure 1: Protein gel data </strong><br />\n CUTANA pAG-Tn5 for CUT&Tag (1 µg) was resolved via SDS-PAGE\n and stained with Coomassie blue. The migration and molecular\n weight of the protein standards are indicated. Uncharged\n pAG-Tn5 monomer is 78.5 kDa, however once charged with DNA,\n Tn5 dimerizes to a final complex weight of 191 kDa.\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/proteins/15-1017-size-distribution.jpeg\"\n target=\"blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"15-1017-size-distribution\"\n src=\"/content/images/products/proteins/15-1017-size-distribution.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n ></a\n >\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\"\n ><strong\n >Figure 2: Size distribution of released chromatin</strong\n ><br />\n CUT&Tag was performed as described above. Recovered DNA was\n directly PCR amplified to produce sequence-ready libraries.\n Agilent TapeStation traces for libraries derived from\n negative control Rabbit IgG (top) and H3K27me3 (bottom) are\n shown. Excised DNA is highly enriched for mononucleosomes\n (peak at ~300 bp reflects ~150 bp insert size).\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/proteins/15-1017-cut-tag-data.jpeg\"\n target=\"blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"15-1017-cut-tag-data\"\n src=\"/content/images/products/proteins/15-1017-cut-tag-data.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n >\n </a>\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\">\n <strong>Figure 3: CUT&Tag data </strong><br />\n CUT&Tag was performed as described above. Representative\n sequencing tracks obtained using CUTANA pAG-Tn5 show a 232\n kb close up view of the LAMC3 gene. CUTANA pAG-Tn5 produced\n clear peaks with genomic distribution profiles consistent\n with the known biological functions of H3K4me3 and H3K27me3\n as well as minimal background in the IgG negative control.\n </span>\n </p>\n </div>\n </div>\n <aside class=\"image-picker__right\">\n <div class=\"image-picker__gallery\">\n <img\n alt=\"15-1017-protein-gel-data\"\n src=\"/content/images/products/proteins/15-1017-protein-gel-data.jpeg\"\n width=\"200\"\n class=\"image-picker__side-image image-picker__side-image_active\"\n role=\"button\"\n />\n <img\n alt=\"15-1017-size-distribution\"\n src=\"/content/images/products/proteins/15-1017-size-distribution.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"15-1017 C&T\"\n src=\"/content/images/products/proteins/15-1017-cut-tag-data.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n </div>\n </aside>\n </section>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Recommended Accessory Reagents</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <table style=\"color: #525051\">\n <tr>\n <th\n style=\"\n padding: 0;\n text-align: left;\n color: #525051;\n text-transform: none;\n \"\n >\n <u>Item</u>\n </th>\n <th style=\"padding: 0; text-align: left; color: #525051\">\n <u>CAT</u>\n </th>\n </tr>\n\n <tr>\n <td>CUTANA™ Concanavalin A Conjugated Paramagnetic Beadss</td>\n <td>\n <a\n href=\"/products/epigenetics-kits-and-reagents/cutana-chic-cut-run-assays/cutana-concanavalin-a-conjugated-paramagnetic-beads\"\n >21-1401</a\n >\n </td>\n </tr>\n\n <tr>\n <td>SNAP-CUTANA™ K-MetStat Panel</td>\n <td>\n <a href=\"/products/nucleosomes/snap-cutana-k-metstat-panel\"\n >19-1002</a\n >\n </td>\n </tr>\n\n <tr>\n <td>Histone H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN</td>\n <td>\n <a\n href=\"/products/antibodies/snap-chip-certified-antibodies/histone-h3k4me3-antibody-snap-chip-certified-cutana-cut-run-compatible\"\n >13-0041</a\n >\n </td>\n </tr>\n\n <tr>\n <td>CUTANA™ Rabbit IgG CUT&RUN Negative Control Antibody</td>\n <td>\n <a\n href=\"/products/nucleosomes/snap-cutana-spike-in-controls/cutana-rabbit-igg-cut-run-negative-control-antibody\"\n >13-0042</a\n >\n </td>\n </tr>\n\n <tr>\n <td>Anti-Rabbit Secondary Antibody for CUTANA™ CUT&Tag</td>\n <td>\n <a\n href=\"/products/epigenetics-reagents-and-assays/cutana-cut-tag-assays/anti-rabbit-secondary-antibody-for-cutana-cut-tag-workflows\"\n >13-0047</a\n >\n </td>\n </tr>\n\n <tr>\n <td>Anti-Mouse Secondary Antibody for CUTANA™ CUT&Tag</td>\n <td>\n <a\n href=\"/products/epigenetics-reagents-and-assays/cutanac-cut-tag-assays/anti-mouse-secondary-antibody-for-cutana-cut-tag-workflows\"\n >13-0048</a\n >\n </td>\n </tr>\n\n <tr>\n <td>CUTANA™ Non-Hot Start 2X PCR Master Mix for CUT&Tag</td>\n <td>\n <a\n href=\"/products/epigenetics-reagents-and-assays/cutana-chic-cut-tag-assays/cutana-high-fidelity-2x-pcr-master-mix\"\n >15-1018</a\n >\n </td>\n </tr>\n\n <tr>\n <td>Magnetic Separation Rack, 0.2 mL Tubes</td>\n <td>\n <a\n href=\"/products/epigenetics-reagents-and-assays/cutana-chic-cut-run-assays/magnetic-separation-rack-0-2-ml-tubes\"\n >10-0008</a\n >\n </td>\n </tr>\n\n <tr>\n <td>Magnetic Separation Rack, 1.5 mL Tubes</td>\n <td>\n <a\n href=\"/products/epigenetics-reagents-and-assays/magnetic-separation-rack-1-5-ml-tubes\"\n >10-0012</a\n >\n </td>\n </tr>\n\n <tr>\n <td>CUTANA™ CUT&RUN 8-strip 0.2 mL Tubes</td>\n <td>\n <a\n href=\"/products/epigenetics-reagents-and-assays/cutana-chic-cut-run-assays/cutana-cut-run-8-strip-0-2-ml-tubes\"\n >10-0009</a\n >\n </td>\n </tr>\n </table>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Technical Information</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <div class=\"product-tech-info\">\n <div class=\"product-tech-info__line-item\">\n <div class=\"product-tech-info__line-item-left\">\n <b>Storage</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n Stable for one year at -20°C from date of receipt. The protein\n is not subject to freeze/thaw under these conditions.\n </div>\n </div>\n <div class=\"product-tech-info__line-item\">\n <div class=\"product-tech-info__line-item-left\">\n <b>Formulation</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n 50 mM HEPES-KOH pH 7.2, 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1%\n Triton X-100, 50% glycerol\n </div>\n </div>\n <div class=\"product-tech-info__line-item\">\n <div class=\"product-tech-info__line-item-left\">\n <b>Adapters</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n <p style=\"margin-bottom: 0\">\n Tn5ME-A: 5’-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3’\n </p>\n <p style=\"margin-bottom: 0\">\n Tn5ME-B: 5’-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-3’\n </p>\n <p style=\"margin-bottom: 0\">\n Tn5ME-rev: 5’-[phos]CTGTCTCTTATACACATCT-3’\n </p>\n </div>\n </div>\n </div>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Application Notes</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <p>\n Add 2.5 µL of the supplied enzyme to a 50 µL CUT&Tag reaction (20X\n dilution). For detailed applications and uses of this product,\n please see our\n <a\n href=\"https://www.epicypher.com/content/documents/protocols/cutana-cut&tag-protocol.pdf\"\n >CUT&Tag protocol</a\n >.\n </p>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">References</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <b>Background References:</b><br />\n [1] Kaya-Okur et al. <em>Nat. Commun.</em> (2019). PMID:\n <a\n href=\"https://pubmed.ncbi.nlm.nih.gov/31036827/\"\n title=\"CUT&Tag for efficient epigenomic profiling of small samples and single cells\"\n target=\"new\"\n >31036827</a\n ><br />\n <br />\n <strong>Product References:</strong>\n <div>\n <iframe\n class=\"airtable-embed\"\n src=\"https://airtable.com/embed/shr4aG2p59KazCtSj?backgroundColor=purple&viewControls=on\"\n frameborder=\"0\"\n onmousewheel=\"\"\n width=\"100%\"\n height=\"300\"\n style=\"background: transparent; border: 1px solid #ccc\"\n ></iframe>\n </div>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Documents & Resources</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <div class=\"product-documents\">\n <a\n href=\"/content/documents/tds/15-1017.pdf\"\n target=\"blank\"\n class=\"product-documents__link\"\n >\n <svg\n version=\"1.1\"\n id=\"Layer_1\"\n xmlns=\"http://www.w3.org/2000/svg\"\n xmlns:xlink=\"http://www.w3.org/1999/xlink\"\n x=\"0px\"\n y=\"0px\"\n viewBox=\"0 0 228 240\"\n style=\"enable-background: new 0 0 228 240\"\n xml:space=\"preserve\"\n class=\"product-documents__icon\"\n >\n <g>\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M191.92,68.77l-47.69-47.69c-1.33-1.33-3.12-2.08-5.01-2.08H45.09C41.17,19,38,22.17,38,26.09v184.36\n c0,3.92,3.17,7.09,7.09,7.09h141.82c3.92,0,7.09-3.17,7.09-7.09V73.8C194,71.92,193.25,70.1,191.92,68.77z M177.65,77.06h-41.7\n v-41.7L177.65,77.06z M178.05,201.59H53.95V34.95h66.92v47.86c0,5.14,4.17,9.31,9.31,9.31h47.86V201.59z\"\n />\n </g>\n <rect\n x=\"20\"\n y=\"112\"\n class=\"product-documents__svg-background\"\n width=\"146\"\n height=\"76\"\n />\n <g>\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M23.83,125.68h22.36c5.29,0,9.41,1.33,12.35,4c2.94,2.67,4.42,6.39,4.42,11.18c0,4.78-1.47,8.51-4.42,11.18\n c-2.94,2.67-7.06,4-12.35,4H34.59v18.29H23.83V125.68z M44.81,147.9c5.38,0,8.07-2.32,8.07-6.97c0-2.39-0.67-4.16-2-5.31\n c-1.33-1.15-3.36-1.73-6.07-1.73H34.59v14.01H44.81z\"\n />\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M69.92,125.68h18.91c5.29,0,9.84,0.97,13.66,2.9c3.82,1.93,6.74,4.72,8.76,8.35\n c2.02,3.63,3.04,7.98,3.04,13.04c0,5.06-1,9.42-3,13.08c-2,3.66-4.91,6.45-8.73,8.38c-3.82,1.93-8.4,2.9-13.73,2.9H69.92V125.68z\n M88.07,165.63c10.35,0,15.52-5.22,15.52-15.66c0-10.4-5.17-15.59-15.52-15.59h-7.38v31.26H88.07z\"\n />\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M122.57,125.68h32.84v8.49h-22.22v11.18h20.84v8.49h-20.84v20.49h-10.63V125.68z\"\n />\n </g>\n </svg>\n <span class=\"product-documents__info\">Technical Datasheet</span>\n </a>\n </div>\n <div class=\"product-documents\">\n <a\n href=\"/content/documents/protocols/cutana-cut&tag-protocol.pdf\"\n target=\"blank\"\n class=\"product-documents__link\"\n >\n <svg\n version=\"1.1\"\n id=\"Layer_1\"\n xmlns=\"http://www.w3.org/2000/svg\"\n xmlns:xlink=\"http://www.w3.org/1999/xlink\"\n x=\"0px\"\n y=\"0px\"\n viewBox=\"0 0 228 240\"\n style=\"enable-background: new 0 0 228 240\"\n xml:space=\"preserve\"\n class=\"product-documents__icon\"\n >\n <g>\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M191.92,68.77l-47.69-47.69c-1.33-1.33-3.12-2.08-5.01-2.08H45.09C41.17,19,38,22.17,38,26.09v184.36\n c0,3.92,3.17,7.09,7.09,7.09h141.82c3.92,0,7.09-3.17,7.09-7.09V73.8C194,71.92,193.25,70.1,191.92,68.77z M177.65,77.06h-41.7\n v-41.7L177.65,77.06z M178.05,201.59H53.95V34.95h66.92v47.86c0,5.14,4.17,9.31,9.31,9.31h47.86V201.59z\"\n />\n </g>\n <rect\n x=\"20\"\n y=\"112\"\n class=\"product-documents__svg-background\"\n width=\"146\"\n height=\"76\"\n />\n <g>\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M23.83,125.68h22.36c5.29,0,9.41,1.33,12.35,4c2.94,2.67,4.42,6.39,4.42,11.18c0,4.78-1.47,8.51-4.42,11.18\n c-2.94,2.67-7.06,4-12.35,4H34.59v18.29H23.83V125.68z M44.81,147.9c5.38,0,8.07-2.32,8.07-6.97c0-2.39-0.67-4.16-2-5.31\n c-1.33-1.15-3.36-1.73-6.07-1.73H34.59v14.01H44.81z\"\n />\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M69.92,125.68h18.91c5.29,0,9.84,0.97,13.66,2.9c3.82,1.93,6.74,4.72,8.76,8.35\n c2.02,3.63,3.04,7.98,3.04,13.04c0,5.06-1,9.42-3,13.08c-2,3.66-4.91,6.45-8.73,8.38c-3.82,1.93-8.4,2.9-13.73,2.9H69.92V125.68z\n M88.07,165.63c10.35,0,15.52-5.22,15.52-15.66c0-10.4-5.17-15.59-15.52-15.59h-7.38v31.26H88.07z\"\n />\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M122.57,125.68h32.84v8.49h-22.22v11.18h20.84v8.49h-20.84v20.49h-10.63V125.68z\"\n />\n </g>\n </svg>\n <span class=\"product-documents__info\"\n >CUT&Tag Protocol for Histone PTMs</span\n >\n </a>\n </div>\n <div class=\"product-documents\">\n <a\n href=\"/resources/protocols/cutana-cut-and-tag-protocol/\"\n target=\"blank\"\n class=\"product-documents__link\"\n >\n <svg\n version=\"1.1\"\n id=\"Layer_1\"\n xmlns=\"http://www.w3.org/2000/svg\"\n xmlns:xlink=\"http://www.w3.org/1999/xlink\"\n x=\"0px\"\n y=\"0px\"\n viewBox=\"0 0 228 240\"\n style=\"enable-background: new 0 0 228 240\"\n xml:space=\"preserve\"\n class=\"product-documents__icon\"\n >\n <g>\n <path\n class=\"product-documents__svg-link\"\n d=\"M130.3,157.06c-0.77-0.76-2-0.76-2.77,0l-28.45,28.45c-13.17,13.17-35.41,14.57-49.95,0\n c-14.57-14.57-13.17-36.78,0-49.95l28.45-28.45c0.76-0.76,0.76-2.01,0-2.77l-9.75-9.75c-0.77-0.76-2-0.76-2.77,0l-28.45,28.45\n c-20.71,20.71-20.71,54.23,0,74.92s54.23,20.71,74.92,0L140,169.52c0.76-0.76,0.76-2.01,0-2.77L130.3,157.06z M193.62,41\n c-20.71-20.71-54.23-20.71-74.92,0L90.22,69.45c-0.76,0.77-0.76,2,0,2.77l9.72,9.72c0.76,0.76,2.01,0.76,2.77,0l28.45-28.45\n c13.17-13.17,35.41-14.57,49.95,0c14.57,14.57,13.17,36.78,0,49.95l-28.45,28.45c-0.76,0.77-0.76,2,0,2.77l9.75,9.75\n c0.76,0.76,2.01,0.76,2.77,0l28.45-28.45C214.31,95.23,214.31,61.71,193.62,41L193.62,41z M139.14,85.29c-0.77-0.76-2-0.76-2.77,0\n l-55.46,55.43c-0.76,0.77-0.76,2,0,2.77l9.7,9.7c0.76,0.76,2.01,0.76,2.77,0l55.43-55.43c0.76-0.76,0.76-2.01,0-2.77L139.14,85.29z\n \"\n />\n </g>\n </svg>\n <span class=\"product-documents__info\"\n >Previous CUTANA<sup>™</sup> CUT&Tag Protocol versions</span\n >\n </a>\n </div>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Intellectual Property</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <p>\n US Pat. No. 10,689,643, 11,306,307, EU Pat. No. 3,688,157, 2,999,784\n and related patents and pending applications\n </p>\n </div>\n </div>\n </div>\n </div>\n</div>\n\n<style>\n table {\n width: 65%;\n }\n th {\n text-decoration: underline;\n }\n td,\n th {\n text-align: left;\n padding: 8px;\n }\n th,\n td,\n tr {\n color: #000;\n }\n li {\n margin-bottom: 0 !important;\n }\n /* Tablet */\n\n @media only screen and (max-width: 949px),\n (min-device-width: 468px) and (max-device-width: 949px) {\n /* Force table to not be like tables anymore */\n table,\n thead,\n tbody,\n th,\n td,\n tr {\n display: block;\n }\n th {\n display: none !important;\n }\n\n /* Hide table headers (but not display: none;, for accessibility) */\n thead tr {\n position: absolute;\n top: -9999px;\n left: -9999px;\n }\n\n tr {\n border: 1px solid #ccc;\n margin-bottom: 1rem !important;\n }\n\n td {\n /* Behave like a \"row\" */\n border: none;\n /* border-bottom: 1px solid lightgray !important; */\n position: relative;\n padding-left: 50%;\n display: 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- Type: Transposase
- Mol Wgt: 191 kDa
- Host: E. coli
- Epitope Tag: None
Description
Available in 50 and 250 Reaction pack sizes. CUTANA pAG-Tn5 is the key reagent for performing efficient mapping of chromatin features in Cleavage Under Targets and Tagmentation (CUT&Tag) [1]. CUT&Tag offers significant improvements in signal to noise at reduced cell inputs and sequencing depth compared to ChIP-seq. As a fusion of Proteins A and G to a highly active E. coli transposase mutant (Tn5), CUTANA pAG-Tn5 is compatible with target antibodies from a broad spectrum of host species. It also comes without an epitope tag, making it suitable for tag-mediated CUT&Tag (e.g. FLAG, HA, TY1, V5, etc.). This product is highly purified to remove contaminating E. coli DNA, which can be tagmented during storage and complicate analysis at low cell numbers. For superior normalization as well as antibody validation and reaction monitoring, SNAP-CUTANA™ nucleosome spike-ins are recommended. The Tn5 comes charged with mosaic adapters and is ready to be used in CUT&Tag.
Validation Data
CUT&Tag was performed on 100k K562 cells with 0.5 µg of either IgG (EpiCypher 13-0042), H3K4me3 (EpiCypher 13-0041), or H3K27me3 (ThermoFisher MA5-11198) antibodies using CUTANA™ pAG-Tn5 (1:20 dilution) following the EpiCypher Direct-to-PCR CUT&Tag protocol. Libraries were run on an Illumina NextSeq2000 with paired-end sequencing (2x50 bp). Sample sequencing depth was 0.3 million reads (IgG), 1.7 million reads (H3K4me3), and 3.8 million reads (H3K27me3). Data were aligned to the hg19 genome using Bowtie2. Data were filtered to remove duplicates, multi-aligned reads, and blacklist regions.
Figure 1: Protein gel data
CUTANA pAG-Tn5 for CUT&Tag (1 µg) was resolved via SDS-PAGE
and stained with Coomassie blue. The migration and molecular
weight of the protein standards are indicated. Uncharged
pAG-Tn5 monomer is 78.5 kDa, however once charged with DNA,
Tn5 dimerizes to a final complex weight of 191 kDa.
Figure 2: Size distribution of released chromatin
CUT&Tag was performed as described above. Recovered DNA was
directly PCR amplified to produce sequence-ready libraries.
Agilent TapeStation traces for libraries derived from
negative control Rabbit IgG (top) and H3K27me3 (bottom) are
shown. Excised DNA is highly enriched for mononucleosomes
(peak at ~300 bp reflects ~150 bp insert size).
Figure 3: CUT&Tag data
CUT&Tag was performed as described above. Representative
sequencing tracks obtained using CUTANA pAG-Tn5 show a 232
kb close up view of the LAMC3 gene. CUTANA pAG-Tn5 produced
clear peaks with genomic distribution profiles consistent
with the known biological functions of H3K4me3 and H3K27me3
as well as minimal background in the IgG negative control.
Recommended Accessory Reagents
Item | CAT |
---|---|
CUTANA™ Concanavalin A Conjugated Paramagnetic Beadss | 21-1401 |
SNAP-CUTANA™ K-MetStat Panel | 19-1002 |
Histone H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN | 13-0041 |
CUTANA™ Rabbit IgG CUT&RUN Negative Control Antibody | 13-0042 |
Anti-Rabbit Secondary Antibody for CUTANA™ CUT&Tag | 13-0047 |
Anti-Mouse Secondary Antibody for CUTANA™ CUT&Tag | 13-0048 |
CUTANA™ Non-Hot Start 2X PCR Master Mix for CUT&Tag | 15-1018 |
Magnetic Separation Rack, 0.2 mL Tubes | 10-0008 |
Magnetic Separation Rack, 1.5 mL Tubes | 10-0012 |
CUTANA™ CUT&RUN 8-strip 0.2 mL Tubes | 10-0009 |
Technical Information
Tn5ME-A: 5’-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3’
Tn5ME-B: 5’-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-3’
Tn5ME-rev: 5’-[phos]CTGTCTCTTATACACATCT-3’
Application Notes
Add 2.5 µL of the supplied enzyme to a 50 µL CUT&Tag reaction (20X dilution). For detailed applications and uses of this product, please see our CUT&Tag protocol.
References
Documents & Resources
Intellectual Property
US Pat. No. 10,689,643, 11,306,307, EU Pat. No. 3,688,157, 2,999,784 and related patents and pending applications
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