{"id":152,"date":"2016-11-02T17:26:41","date_gmt":"2016-11-02T17:26:41","guid":{"rendered":"https:\/\/umaine.edu\/dnaseq\/?page_id=152"},"modified":"2025-06-03T10:45:59","modified_gmt":"2025-06-03T14:45:59","slug":"prepare-your-template","status":"publish","type":"page","link":"https:\/\/umaine.edu\/core\/dna-sequencing\/ordering\/prepare-your-template\/","title":{"rendered":"1. Prepare your template"},"content":{"rendered":"\n<div class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-constrained wp-block-group-is-layout-constrained\">\n<h3 class=\"wp-block-heading\"><strong>Please prepare your template using our checklist:<\/strong><\/h3>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Ensure that any kits you use to prepare samples specify that DNA sequencing is a downstream application.<\/li>\n\n\n\n<li>Submit DNA templates in either sterile water or 10mM Tris pH 8.0.<\/li>\n\n\n\n<li>Supply 10 uL of DNA per reaction. Only 1 uL of DNA is used per reaction but providing additional volume allows for re-runs if necessary. If sample volume is a concern, please discuss with the Lab Coordinator.<\/li>\n\n\n\n<li>Review the following table for recommended quantity measurements. Final concentration will be in units of ng\/uL or ug\/uL depending on DNA template.<\/li>\n<\/ul>\n\n\n<style>.wp-block-kadence-spacer.kt-block-spacer-152_1c317b-ec .kt-block-spacer{height:20px;}.wp-block-kadence-spacer.kt-block-spacer-152_1c317b-ec .kt-divider{border-top-width:1px;height:1px;border-top-color:#eee;width:80%;border-top-style:solid;}<\/style>\n<div class=\"wp-block-kadence-spacer aligncenter kt-block-spacer-152_1c317b-ec\"><div class=\"kt-block-spacer kt-block-spacer-halign-center\"><\/div><\/div>\n<\/div><\/div>\n\n\n\n<figure class=\"wp-block-table is-style-stripes\"><table class=\"has-fixed-layout\"><thead><tr><th class=\"has-text-align-center\" data-align=\"center\">Template DNAs<\/th><th class=\"has-text-align-center\" data-align=\"center\">Quantity per reaction<\/th><\/tr><\/thead><tbody><tr><td class=\"has-text-align-center\" data-align=\"center\">100-200 bp<\/td><td class=\"has-text-align-center\" data-align=\"center\">1-3 ng<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">200-500 bp<\/td><td class=\"has-text-align-center\" data-align=\"center\">3-10 ng<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">500-1000 bp<\/td><td class=\"has-text-align-center\" data-align=\"center\">5-10 ng<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">1000-2000 bp<\/td><td class=\"has-text-align-center\" data-align=\"center\">10-40 ng<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">more than 2000 bp<\/td><td class=\"has-text-align-center\" data-align=\"center\">25-50 ng<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">Single-stranded DNA<\/td><td class=\"has-text-align-center\" data-align=\"center\">25-50 ng<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">Double-stranded DNA<\/td><td class=\"has-text-align-center\" data-align=\"center\">150-300 ng<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">Cosmid, BAC<\/td><td class=\"has-text-align-center\" data-align=\"center\">0.5-1.0 \u00b5g<\/td><\/tr><tr><td class=\"has-text-align-center\" data-align=\"center\">Bacterial genomic DNA<\/td><td class=\"has-text-align-center\" data-align=\"center\">0.5-1.0 \u00b5g<\/td><\/tr><\/tbody><\/table><\/figure>\n\n\n<style>.wp-block-kadence-spacer.kt-block-spacer-152_0c8db4-1d .kt-block-spacer{height:20px;}.wp-block-kadence-spacer.kt-block-spacer-152_0c8db4-1d .kt-divider{border-top-width:1px;height:1px;border-top-color:#eee;width:80%;border-top-style:solid;}<\/style>\n<div class=\"wp-block-kadence-spacer aligncenter kt-block-spacer-152_0c8db4-1d\"><div class=\"kt-block-spacer kt-block-spacer-halign-center\"><\/div><\/div>\n\n\n\n<div class=\"wp-block-group\"><div class=\"wp-block-group__inner-container is-layout-constrained wp-block-group-is-layout-constrained\">\n<h3 class=\"wp-block-heading\"><strong>Fragment Analysis<\/strong><\/h3>\n\n\n\n<p>Fragment analysis plates should be fully prepared by the client and will be run as is.<\/p>\n<\/div><\/div>\n","protected":false},"excerpt":{"rendered":"<p>Please prepare your template using our checklist: Template DNAs Quantity per reaction 100-200 bp 1-3 ng 200-500 bp 3-10 ng 500-1000 bp 5-10 ng 1000-2000 bp 10-40 ng more than 2000 bp 25-50 ng Single-stranded DNA 25-50 ng Double-stranded DNA 150-300 ng Cosmid, BAC 0.5-1.0 \u00b5g Bacterial genomic DNA 0.5-1.0 \u00b5g Fragment Analysis Fragment analysis [&hellip;]<\/p>\n","protected":false},"author":2276,"featured_media":0,"parent":2400,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"templates\/page-withsidebar.php","meta":{"_seopress_robots_primary_cat":"","_seopress_titles_title":"","_seopress_titles_desc":"","_seopress_robots_index":"","_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0,"footnotes":""},"class_list":["post-152","page","type-page","status-publish","hentry"],"taxonomy_info":[],"featured_image_src_large":false,"author_info":{"display_name":"jrutherford","author_link":"https:\/\/umaine.edu\/core\/author\/jrutherford\/"},"comment_info":0,"_links":{"self":[{"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/pages\/152","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/users\/2276"}],"replies":[{"embeddable":true,"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/comments?post=152"}],"version-history":[{"count":1,"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/pages\/152\/revisions"}],"predecessor-version":[{"id":2448,"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/pages\/152\/revisions\/2448"}],"up":[{"embeddable":true,"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/pages\/2400"}],"wp:attachment":[{"href":"https:\/\/umaine.edu\/core\/wp-json\/wp\/v2\/media?parent=152"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}