{"id":8088,"date":"2016-03-24T13:13:52","date_gmt":"2016-03-24T13:13:52","guid":{"rendered":"http:\/\/admin.acceleratingscience.com\/?p=8088"},"modified":"2020-09-26T00:17:19","modified_gmt":"2020-09-26T00:17:19","slug":"how-to-generate-high-quality-sanger-sequencing-data","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/behindthebench\/how-to-generate-high-quality-sanger-sequencing-data\/","title":{"rendered":"How to Generate High Quality Sanger Sequencing Data"},"content":{"rendered":"<p>Considered as the gold standard, <a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/sequencing\/sanger-sequencing.html?socid=social_btb_abseq\" target=\"_blank\" rel=\"noopener noreferrer\">Sanger sequencing<\/a> technology has been proven through decades of results. You might be familiar with the simple workflow or already have an established protocol in your lab.\u00a0 However, with the new applications developed for Sanger sequencing such as <a href=\"http:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/sequencing\/sanger-sequencing\/sanger-dna-sequencing\/sanger-sequencing-data-analysis\/minor-variant-finder-software.html?socid=social_btb_abseq\" target=\"_blank\" rel=\"noopener noreferrer\">minor variant detection<\/a>, the data quality that you are getting out of your protocol just might not be up to par.<\/p>\n<p>To help researchers generate high-quality data for minor variant detection, we put together a few user bulletins to provide some best practices and troubleshooting tips to help make your life in the lab easier.\u00a0 These protocols have been extensively tested to ensure high quality, reproducible results. Check it out!\u00a0 You might uncover a new possibility for your <a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/sequencing\/sanger-sequencing\/sanger-sequencing-technology-accessories.html?socid=social_btb_abseq\" target=\"_blank\" rel=\"noopener noreferrer\">Capillary Electrophoresis instrument<\/a>\u00a0and perhaps finally resolve that data issue that you\u2019ve been trying to live with for years.<\/p>\n<h3><a style=\"line-height: 1.5\" href=\"http:\/\/tools.thermofisher.com\/content\/sfs\/manuals\/MAN0014628_BigDyeTermin_Cycle_Seq_hiqual_data_UB.pdf?cid=techcomm94?socid=social_btb_abseq\" target=\"_blank\" rel=\"noopener noreferrer\"><strong>Generating high\u2011quality data using the BigDye \u2122 Terminator v3.1 Cycle Sequencing Kit<\/strong><\/a><\/h3>\n<div class=\"mceTemp\"><\/div>\n<p><a href=\"http:\/\/tools.thermofisher.com\/content\/sfs\/manuals\/MAN0014628_BigDyeTermin_Cycle_Seq_hiqual_data_UB.pdf?cid=techcomm94?socid=social_btb_abseq\"><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-8091\" src=\"http:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech1-232x300.jpg\" alt=\"tech1\" width=\"107\" height=\"139\" srcset=\"https:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech1-232x300.jpg 232w, https:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech1.jpg 265w\" sizes=\"auto, (max-width: 107px) 100vw, 107px\" \/><\/a><\/p>\n<p>A protocol for generating high\u2011quality data for use in minor variant detection using:<\/p>\n<ul>\n<li><a href=\"https:\/\/www.thermofisher.com\/order\/catalog\/product\/4337458?socid=social_btb_abseq\" target=\"_blank\" rel=\"noopener noreferrer\">BigDye\u2122 Terminator v3.1 Cycle Sequencing Kit<\/a><\/li>\n<li>Applied Biosystems\u2122 Genetic Analyzers with POP\u20116\u2122 or POP\u20117\u2122 polymer<\/li>\n<\/ul>\n<h3><a href=\"http:\/\/tools.thermofisher.com\/content\/sfs\/manuals\/MAN0014436_BigDyeDirect_Cycle_Seq_hiqual_data_UB.pdf?cid=techcomm93?socid=social_btb_abseq\" target=\"_blank\" rel=\"noopener noreferrer\"><strong>Generating high\u2011quality data using the BigDye \u2122 Direct Cycle Sequencing Kit<\/strong><\/a><\/h3>\n<p><a href=\"http:\/\/tools.thermofisher.com\/content\/sfs\/manuals\/MAN0014436_BigDyeDirect_Cycle_Seq_hiqual_data_UB.pdf?cid=techcomm93?socid=social_btb_abseq\" target=\"_blank\" rel=\"attachment wp-att-8090 noopener noreferrer\"><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-8090 \" src=\"http:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech2-233x300.jpg\" alt=\"tech2\" width=\"107\" height=\"138\" srcset=\"https:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech2-233x300.jpg 233w, https:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech2.jpg 264w\" sizes=\"auto, (max-width: 107px) 100vw, 107px\" \/><\/a><\/p>\n<p>Sequencing with the<a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/sequencing\/sanger-sequencing\/sanger-sequencing-kits-reagents\/bigdye-direct-sanger-sequencing-kit.html?socid=social_btb_abseq\" target=\"_blank\" rel=\"noopener noreferrer\"> BigDye Direct Cycle Sequencing Kit<\/a> takes fewer steps and less hands-on time.\u00a0 By combing PCR clean-up and cycle sequencing into a single step, the BigDye Direct workflow reduces the traditional Sanger sequencing workflow time by up to 40%. This protocol is also optimized for sequencing short amplicons, since BigDye Direct\u2019s novel sequencing primer and chemistry enable high data quality from base 1 after the sequencing primer.<\/p>\n<h3><a href=\"http:\/\/tools.thermofisher.com\/content\/sfs\/manuals\/MAN0014435_Trbleshoot_Sanger_seq_data_UB.pdf?cid=techcomm92?socid=social_btb_abseq\" target=\"_blank\" rel=\"noopener noreferrer\"><strong>Troubleshooting Sanger sequencing data<br \/>\n<\/strong><\/a><\/h3>\n<div class=\"mceTemp\"><\/div>\n<p><a href=\"http:\/\/tools.thermofisher.com\/content\/sfs\/manuals\/MAN0014435_Trbleshoot_Sanger_seq_data_UB.pdf?cid=techcomm92?socid=social_btb_abseq\"><img loading=\"lazy\" decoding=\"async\" class=\"wp-image-8089\" src=\"http:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech3-231x300.jpg\" alt=\"tech3\" width=\"109\" height=\"142\" srcset=\"https:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech3-231x300.jpg 231w, https:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/tech3.jpg 239w\" sizes=\"auto, (max-width: 109px) 100vw, 109px\" \/><\/a><\/p>\n<p>Do you experience signal saturation, low signal intensity, dye blobs\u2026etc. from your data? This document provides guidance with troubleshooting tips for improving sequencing data quality.<\/p>\n<p>Did these help you solve your problems and achieve higher quality data? Let us know what you think about these tips and tricks in the comments section below.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Considered as the gold standard, Sanger sequencing technology has been proven through decades of results. You might be familiar with the simple workflow or already have an established protocol in your lab.\u00a0 However, with the new applications developed for Sanger sequencing such as minor variant detection, the data quality that you are getting out of<\/p>\n","protected":false},"author":120,"featured_media":8403,"comment_status":"open","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":""},"categories":[108],"tags":[17,36,99,153,25,74],"division":[],"class_list":{"0":"post-8088","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-lab-tips-and-tricks","8":"tag-applied-biosystems","9":"tag-capillary-electrophoresis","10":"tag-fragment-analysis","11":"tag-genomic-variants","12":"tag-sanger-sequencing","13":"tag-sequencing","14":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"<!-- This site is optimized with the Yoast SEO Premium plugin v27.8 (Yoast SEO v27.8) - https:\/\/yoast.com\/product\/yoast-seo-premium-wordpress\/ -->\n<title>How to Generate High Quality Sanger Sequencing Data - Behind the Bench<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www.thermofisher.com\/blog\/behindthebench\/how-to-generate-high-quality-sanger-sequencing-data\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"How to Generate High Quality Sanger Sequencing Data\" \/>\n<meta property=\"og:description\" content=\"Considered as the gold standard, Sanger sequencing technology has been proven through decades of results. You might be familiar with the simple workflow or already have an established protocol in your lab.\u00a0 However, with the new applications developed for Sanger sequencing such as minor variant detection, the data quality that you are getting out of\" \/>\n<meta property=\"og:url\" content=\"https:\/\/www.thermofisher.com\/blog\/behindthebench\/how-to-generate-high-quality-sanger-sequencing-data\/\" \/>\n<meta property=\"og:site_name\" content=\"Behind the Bench\" \/>\n<meta property=\"article:publisher\" content=\"https:\/\/www.facebook.com\/thermofisher\" \/>\n<meta property=\"article:published_time\" content=\"2016-03-24T13:13:52+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2020-09-26T00:17:19+00:00\" \/>\n<meta property=\"og:image\" content=\"https:\/\/admin.acceleratingscience.com\/behindthebench\/wp-content\/uploads\/sites\/9\/2016\/03\/Digital-illustration-of-a-DNA-strand.jpg\" \/>\n\t<meta property=\"og:image:width\" content=\"1000\" \/>\n\t<meta property=\"og:image:height\" content=\"667\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/jpeg\" \/>\n<meta name=\"author\" content=\"Behind The Bench Staff\" \/>\n<meta name=\"twitter:card\" content=\"summary_large_image\" \/>\n<meta name=\"twitter:creator\" content=\"@thermofisher\" \/>\n<meta name=\"twitter:site\" content=\"@thermofisher\" \/>\n<meta name=\"twitter:label1\" content=\"Written by\" \/>\n\t<meta name=\"twitter:data1\" content=\"Behind The Bench Staff\" \/>\n\t<meta name=\"twitter:label2\" content=\"Est. reading time\" \/>\n\t<meta name=\"twitter:data2\" content=\"2 minutes\" \/>\n<script type=\"application\/ld+json\" class=\"yoast-schema-graph\">{\"@context\":\"https:\\\/\\\/schema.org\",\"@graph\":[{\"@type\":\"Article\",\"@id\":\"https:\\\/\\\/www.thermofisher.com\\\/blog\\\/behindthebench\\\/how-to-generate-high-quality-sanger-sequencing-data\\\/#article\",\"isPartOf\":{\"@id\":\"https:\\\/\\\/www.thermofisher.com\\\/blog\\\/behindthebench\\\/how-to-generate-high-quality-sanger-sequencing-data\\\/\"},\"author\":{\"name\":\"Behind The Bench Staff\",\"@id\":\"https:\\\/\\\/admin.acceleratingscience.com\\\/behindthebench\\\/#\\\/schema\\\/person\\\/9bbb2d67616274d3d4a21a584f7f4a0c\"},\"headline\":\"How to Generate High Quality Sanger Sequencing Data\",\"datePublished\":\"2016-03-24T13:13:52+00:00\",\"dateModified\":\"2020-09-26T00:17:19+00:00\",\"mainEntityOfPage\":{\"@id\":\"https:\\\/\\\/www.thermofisher.com\\\/blog\\\/behindthebench\\\/how-to-generate-high-quality-sanger-sequencing-data\\\/\"},\"wordCount\":327,\"commentCount\":0,\"image\":{\"@id\":\"https:\\\/\\\/www.thermofisher.com\\\/blog\\\/behindthebench\\\/how-to-generate-high-quality-sanger-sequencing-data\\\/#primaryimage\"},\"thumbnailUrl\":\"https:\\\/\\\/admin.acceleratingscience.com\\\/behindthebench\\\/wp-content\\\/uploads\\\/sites\\\/9\\\/2016\\\/03\\\/Digital-illustration-of-a-DNA-strand.jpg\",\"keywords\":[\"Applied Biosystems\",\"Capillary Electrophoresis\",\"Fragment Analysis\",\"Genomic Variants\",\"Sanger Sequencing\",\"sequencing\"],\"articleSection\":[\"Lab Tips and Tricks\"],\"inLanguage\":\"en-US\",\"potentialAction\":[{\"@type\":\"CommentAction\",\"name\":\"Comment\",\"target\":[\"https:\\\/\\\/www.thermofisher.com\\\/blog\\\/behindthebench\\\/how-to-generate-high-quality-sanger-sequencing-data\\\/#respond\"]}]},{\"@type\":\"WebPage\",\"@id\":\"https:\\\/\\\/www.thermofisher.com\\\/blog\\\/behindthebench\\\/how-to-generate-high-quality-sanger-sequencing-data\\\/\",\"url\":\"https:\\\/\\\/www.thermofisher.com\\\/blog\\\/behindthebench\\\/how-to-generate-high-quality-sanger-sequencing-data\\\/\",\"name\":\"How to Generate High Quality Sanger Sequencing Data - 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