

CUTANA™ E. coli Spike-in DNA
{"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=748","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™ ChIC / CUT&RUN Assays"],"custom_fields":[{"id":"510","name":"Pack size","value":"100 ng"}],"customizations":[],"description":"<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 >\n <p>\n Fragmented DNA derived from <em>Escherichia coli (E. coli)</em> can\n be used as a spike-in control for experimental normalization in\n Cleavage Under Targets and Release Using Nuclease (CUT&RUN). CUT&RUN\n normalization was originally performed with carryover\n <em>E. coli</em> DNA from MNase preparation [1], however pure preps\n of enzyme require post hoc DNA spike-in. CUTANA™\n <em>E. coli</em> Spike-in DNA contains sufficient material for\n 100-200 CUT&RUN samples (range for high and low abundance targets).\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 >\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/nucleosomes/18-1401-dna-gel-data.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"18-1401-dna-gel-data\"\n src=\"/content/images/products/nucleosomes/18-1401-dna-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: DNA gel data </strong><br />\n <em>E. coli</em> fragment size distribution.\n <strong>Lane 1</strong>: gDNA extracted from JM101\n <em>E. coli</em> cells (500 ng). <strong>Lane 2</strong>:\n Digested and purified CUTANA™ <em>E. coli</em> Spike-in DNA\n (500 ng) resolved via 2% E-Gel™ EX Agarose Gel. Migration\n positions of DNA molecular weight markers are indicated.\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/nucleosomes/18-1401-sequencing-data.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"18-1401-sequencing-data\"\n src=\"/content/images/products/nucleosomes/18-1401-sequencing-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 2: CUT&RUN sequencing data</strong><br />\n CUTANA™ <em>E. coli</em> Spike-in DNA (1.0 ng) was added to\n CUT&RUN samples using 500,000 K562 cells enriched for a low\n abundance target (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 >), a high abundance target (H3K27me3, EpiCypher\n <a\n href=\"/products/antibodies/snap-chip-certified-antibodies/histone-h3k27me3-antibody-snap-chip-certified-cutana-cut-run-compatible-discontinued\"\n >13-0030</a\n >) and an IgG negative control (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 >). Total numbers of paired-end sequencing reads, reads\n aligned to <em>E. coli</em>, and percentage of total\n sequencing reads aligned to <em>E. coli</em> spike-in DNA\n are shown.\n <strong\n >NOTE: Spike-in DNA amount should be optimized by the end\n user with the goal of <em>E. coli</em> DNA comprising ~1%\n (0.2 - 5%) of total sequencing reads.</strong\n >\n </span>\n </p>\n </div>\n </div>\n <aside class=\"image-picker__right\">\n <div class=\"image-picker__gallery\">\n <img\n alt=\"18-1401-dna-gel-data\"\n src=\"/content/images/products/nucleosomes/18-1401-dna-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=\"18-1401-sequencing-data\"\n src=\"/content/images/products/nucleosomes/18-1401-sequencing-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\">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>Formulation</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n 100 ng lyophilized DNA.\n <strong>*NOTE</strong>: May not be visible.\n </div>\n </div>\n <div class=\"product-tech-info__line-item\">\n <div class=\"product-tech-info__line-item-left\">\n <b>Storage and Stability</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n Stable for 2 years at -20°C from date of receipt. After\n resuspending, aliquots should be stored at -20°C.\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 Prior to opening, pellet DNA by quickly spinning in a benchtop\n microfuge. Reconstitute in 200 µL DNase free water (0.5 ng/µL).\n Vortex tube on all sides to ensure complete resuspension. Quick spin\n in benchtop microfuge prior to use.\n </p>\n\n <p>\n <strong>Use in CUT&RUN:</strong>\n </p>\n <ol>\n <li>\n Use with CUTANA™ pAG-MNase for ChIC/CUT&RUN (EpiCypher\n <a\n href=\"/products/epigenetics-reagents-and-assays/cutana-pag-mnase-for-chic-cut-and-run-workflows\"\n >15-1016</a\n >), which has very low <em>E. coli</em> DNA carryover.\n </li>\n\n <li>\n Add 1-2 µL* (0.5-1 ng) <em>E. coli</em> Spike-in DNA directly to\n the Stop Buffer, which quenches calcium-mediated pAG-MNase DNA\n digestion. Gently vortex to mix, and add to samples. <br />\n <strong>*NOTE</strong>: Based on target abundance and antibody\n efficiency, the amount of <em>E. coli</em> DNA may need to be\n adjusted. Aim for spike-in DNA to comprise ~1% (0.2-5%) of total\n sequencing reads (<strong>Figure 2</strong>).\n </li>\n <li>\n After sequencing, align reads to the reference genome of\n experimental samples (e.g. human) as well as to\n <em>E. coli</em> genome.\n </li>\n <li>\n Normalize data by following recommendations in the\n <a href=\"/resources/protocols/cutana-cut-and-run-protocol/\"\n >CUT&RUN Protocol</a\n >.\n </li>\n </ol>\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 <strong>Background References:</strong>\n <br />\n [1] Meers et al. <em>Elife.</em> (2019). PMID:\n <a\n href=\"https://pubmed.ncbi.nlm.nih.gov/31232687/\"\n target=\"_blank\"\n title=\"Improved CUT&RUN chromatin profiling tools\"\n >31232687</a\n ><br />\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/18-1401.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 alt=\"16-0030 Datasheet\"\n >\n <g>\n <path\n class=\"product-documents__svg-pdf\"\n 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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>\n </div>\n </div>\n </div>\n</div>\n","detail_messages":"","gift_wrapping_available":false,"gtin":null,"id":"748","images":[{"alt":"CUTANA™ E. coli Spike-in DNA","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/748/730/DNA_Vector_Draw_Final-w-border-01__76823.1592491202.png?c=2"}],"main_image":{"alt":"CUTANA™ E. coli Spike-in 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\n Type: Polyclonal\n \n \n Host: Rabbit\n \n \n Applications: CUT&R","url":"https://www.epicypher.com/products/antibodies/cutana-cut-run-antibodies/cut-run-antibodies-chromatin-associated-proteins/menin-cutana-cut-run-antibody","weight":{"formatted":"0.01 LBS","value":0.01}}],"shipping":{"calculated":true},"shipping_messages":[],"show_quantity_input":1,"sku":"18-1401","tags":[],"title":"CUTANA™ E. coli Spike-in DNA","upc":null,"url":"https://www.epicypher.com/products/epigenetics-reagents-and-assays/cutana-e-coli-spike-in-dna","warranty":"","weight":"0.01 LBS"} Pack size: 100 ng
Description
Fragmented DNA derived from Escherichia coli (E. coli) can be used as a spike-in control for experimental normalization in Cleavage Under Targets and Release Using Nuclease (CUT&RUN). CUT&RUN normalization was originally performed with carryover E. coli DNA from MNase preparation [1], however pure preps of enzyme require post hoc DNA spike-in. CUTANA™ E. coli Spike-in DNA contains sufficient material for 100-200 CUT&RUN samples (range for high and low abundance targets).
Validation Data
Figure 1: DNA gel data
E. coli fragment size distribution.
Lane 1: gDNA extracted from JM101
E. coli cells (500 ng). Lane 2:
Digested and purified CUTANA™ E. coli Spike-in DNA
(500 ng) resolved via 2% E-Gel™ EX Agarose Gel. Migration
positions of DNA molecular weight markers are indicated.
Figure 2: CUT&RUN sequencing data
CUTANA™ E. coli Spike-in DNA (1.0 ng) was added to
CUT&RUN samples using 500,000 K562 cells enriched for a low
abundance target (H3K4me3, EpiCypher
13-0041), a high abundance target (H3K27me3, EpiCypher
13-0030) and an IgG negative control (EpiCypher
13-0042). Total numbers of paired-end sequencing reads, reads
aligned to E. coli, and percentage of total
sequencing reads aligned to E. coli spike-in DNA
are shown.
NOTE: Spike-in DNA amount should be optimized by the end
user with the goal of E. coli DNA comprising ~1%
(0.2 - 5%) of total sequencing reads.
Technical Information
Application Notes
Prior to opening, pellet DNA by quickly spinning in a benchtop microfuge. Reconstitute in 200 µL DNase free water (0.5 ng/µL). Vortex tube on all sides to ensure complete resuspension. Quick spin in benchtop microfuge prior to use.
Use in CUT&RUN:
- Use with CUTANA™ pAG-MNase for ChIC/CUT&RUN (EpiCypher 15-1016), which has very low E. coli DNA carryover.
-
Add 1-2 µL* (0.5-1 ng) E. coli Spike-in DNA directly to
the Stop Buffer, which quenches calcium-mediated pAG-MNase DNA
digestion. Gently vortex to mix, and add to samples.
*NOTE: Based on target abundance and antibody efficiency, the amount of E. coli DNA may need to be adjusted. Aim for spike-in DNA to comprise ~1% (0.2-5%) of total sequencing reads (Figure 2). - After sequencing, align reads to the reference genome of experimental samples (e.g. human) as well as to E. coli genome.
- Normalize data by following recommendations in the CUT&RUN Protocol.
References
Documents & Resources
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