[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-1-en-105":3,"doc-seo-243627-105":53,"doc-detail-243627-en":126},{"code":4,"msg":5,"data":6},0,"success",[7,14,19,24,29,34,39,44,49],{"id":8,"doc_module":9,"doc_module_name":10,"category_name":11,"show_sort_weight":12,"slug":13},11,1,"Template","Presentations",90,"presentations",{"id":15,"doc_module":9,"doc_module_name":10,"category_name":16,"show_sort_weight":17,"slug":18},12,"Resumes",80,"resumes",{"id":20,"doc_module":9,"doc_module_name":10,"category_name":21,"show_sort_weight":22,"slug":23},14,"Invoices",70,"invoices",{"id":25,"doc_module":9,"doc_module_name":10,"category_name":26,"show_sort_weight":27,"slug":28},15,"Posters",60,"posters",{"id":30,"doc_module":9,"doc_module_name":10,"category_name":31,"show_sort_weight":32,"slug":33},16,"Social Media",50,"social-media",{"id":35,"doc_module":9,"doc_module_name":10,"category_name":36,"show_sort_weight":37,"slug":38},17,"Forms",40,"forms",{"id":40,"doc_module":9,"doc_module_name":10,"category_name":41,"show_sort_weight":42,"slug":43},18,"Letters",30,"letters",{"id":45,"doc_module":9,"doc_module_name":10,"category_name":46,"show_sort_weight":47,"slug":48},21,"Paper Templates",5,"papers-templates",{"id":50,"doc_module":9,"doc_module_name":10,"category_name":51,"show_sort_weight":4,"slug":52},158,"General","general-158",{"code":4,"msg":54,"data":55},"ok",{"site_id":56,"language":57,"slug":58,"title":59,"keywords":60,"description":61,"schema_data":62,"social_meta":119,"head_meta":121,"extra_data":123,"updated_unix":125},105,"en","magnetic-bead-purification-of-labeled-dna-fragments-for-high-throughput-capillary-electrophoresis-sequencing","Magnetic Bead Purification of Labeled DNA Fragments for High-Throughput Capillary Electrophoresis Sequencing","","Automated purification of terminator sequencing products is presented using a magnetic bead workflow for 384-well plates. The method yields labeled DNA fragments essentially free of contaminants at very low cost per reaction and streamlines sample handling by avoiding laborious precipitation steps. Compared with ethanol precipitation, it increases phred20 read length by about forty bases across PCR fragments, plasmids, cosmids, and RCA products. It is compatible with MegaBACE 1000 and ABI Prism 3700 instruments and has produced large-scale sequencing output.",{"@graph":63,"@context":118},[64,80,101],{"@type":65,"itemListElement":66},"BreadcrumbList",[67,71,74,77],{"item":68,"name":69,"@type":70,"position":9},"https://docshare.wps.com","Home","ListItem",{"item":72,"name":10,"@type":70,"position":73},"https://docshare.wps.com/template/",2,{"item":75,"name":46,"@type":70,"position":76},"https://docshare.wps.com/template/paper-templates/",3,{"item":78,"name":59,"@type":70,"position":79},"https://docshare.wps.com/template/magnetic-bead-purification-of-labeled-dna-fragments-for-high-throughput-capillary-electrophoresis-sequencing/243627/",4,{"url":78,"name":59,"@type":81,"image":82,"author":87,"headline":59,"publisher":90,"fileFormat":93,"inLanguage":57,"description":61,"dateModified":94,"datePublished":95,"encodingFormat":93,"isAccessibleForFree":96,"interactionStatistic":97},"DigitalDocument",{"url":83,"@type":84,"width":85,"height":86},"https://docshare.wps.com/thumbnails/magnetic-bead-purification-of-labeled-dna-fragments-for-high-throughput-capillary-electrophoresis-sequencing/243627.png","ImageObject",442,249,{"name":88,"@type":89},"Aladdin","Person",{"url":68,"name":91,"@type":92},"DocShare","Organization","application/pdf","2026-10-16","2026-09-12",true,{"@type":98,"interactionType":99,"userInteractionCount":76},"InteractionCounter",{"@type":100},"ViewAction",{"@type":102,"mainEntity":103},"FAQPage",[104,110,114],{"name":105,"@type":106,"acceptedAnswer":107},"What sequencing purification approach is described in the document?","Question",{"text":108,"@type":109},"An automated magnetic bead purification workflow is used to clean terminator sequencing products in a 384-well format.","Answer",{"name":111,"@type":106,"acceptedAnswer":112},"How does the magnetic bead method compare with ethanol precipitation?",{"text":113,"@type":109},"Compared with laborious ethanol precipitation, it increases phred20 read length by about forty bases while producing highly pure labeled DNA fragments.",{"name":115,"@type":106,"acceptedAnswer":116},"Which capillary electrophoresis instruments are compatible with the method?",{"text":117,"@type":109},"The protocol is compatible with both the MegaBACE 1000 and ABI Prism 3700 capillary instruments.","https://schema.org",{"og:url":78,"og:type":120,"og:title":59,"og:site_name":91,"og:description":61},"article",{"robots":122,"canonical":78},"index,follow",{"doc_id":124,"site_id":56},243627,1792088825,{"code":4,"msg":5,"data":127},{"doc_id":124,"user_id":128,"nickname":88,"user_avatar":129,"doc_module":9,"category_id":45,"category_name":46,"doc_title":59,"doc_description":61,"doc_content":130,"file_id":131,"file_url":132,"file_type":133,"file_size":134,"view_count":76,"is_deleted":4,"is_public":9,"is_downloadable":9,"audit_status":9,"page_count":47,"language":135,"language_code":57,"site_id":56,"html_lang":57,"table_of_contents":136,"faqs":137,"seo_title":138,"seo_description":61,"update_tm":139,"read_time":73},2336478503145,"https://ap-avatar.wpscdn.com/davatar_276721f389ce27ea32af1340a28f341c","Magnetic Bead Purification of Labeled DNA Fragments for High-Throughput Capillary Electrophoresis Sequencing  \nABSTRACT  \nWe have developed an automated purification method for terminator sequencing products based on a magnetic bead technology. This 384-well protocol generates labeled DNA fragments that are essentially free of contaminates for less than $0.005 per reaction. In comparison to laborious ethanol precipitation protocols, this method increases the phred20 read length by forty bases with various DNA templates such as PCR fragments, Plasmids, Cosmids and RCA products. Our method eliminatescentrifugation and is compatible with both the MegaBACE 1000 and ABI Prism 3700 capillary instruments. As of September 2001, this method has produced over 1.6 million samples with 93% averaging 620 phred20 bases as part of Joint Genome Institutes Production Process.  \nINTRODUCTION  \nIn the late 1990’s, high-throughput capillary electrophoresis instruments were introduced to meet the demands of large scale sequencing labs working on the Human Genome Project. While these instruments provided faster turnaround times, they also created a need for large numbers of high purity DNA samples. The typical approach was to scale up and automate molecular biology methods designed for small-scale bench work. Since these methods often contained laborious and irreproducible steps like centrifugation, we decided to design a  \nhigh-throughput method tailored for robotic platforms and capillary electrophoresis instruments.  \nMATERIAL AND METHODS  \nSee Table 1 for purification protocol.  \nRESULTS AND DISCUSSION  \nIn developing a high-throughput method to purify DNA sequencing fragments, we began with a magnetic bead technology previously used at our facility to purify template DNA for sequencing(1) . This method bound crude DNA to carboxylated  \nTable 1. Sequencing Fragment Purification Protocol  \n1. Sequence RCA generated template with Amersham Pharmacia ET Terminator Kit (part\\# US81095) according to instructions except; reduce final volume to 5ul and process in a 384 well PCR plate.  \n2. Add 10uL of BET solution to each well with 384-well Multidrop.  \n3. Verify solution is mixed thoroughly. Mix by pipetting or vortex as needed.  \n4. Incubate at room temperature for 15 minutes.  \n5. Place 384 well plate on a plate magnet for 1 minute.  \n6. Place 384 well plate/magnet assembly on 384 Hydra and aspirate solution.  \n7. Add 15ul of 70% ethanol solution to each well with 384 well Multidrop  \n8. Place 384 well plate/magnet assembly on 384 Hydra and aspirate solution.  \n9. Air-dry samples for 10 minutes or continue to step 10.  \n10. Dispense 15uL of deionized water to each plate.  \n11. Mix by pipetting or vortex until beads are resuspended  \n12. Incubate 10 minutes at room temperature.  \n13. Place 384 well plate on plate magnet for 2 minutes.  \n14. Transfer 10ul of water solution to suitable plate for electrokinetic injection  \n15. Cover and store at-20°C or load onto MegaBACE 1000 or ABI 3700  \n16. MegaBACE 1000 (Inject-1.6KV/36 seconds, Run-7KV/170 minutes)  \nReagent/Stock Preparation  \nBET Solution (processes twenty 384-well plates)  \nEthanol(100%) 64.0 mls  \nDeionized Water 7.0 mls  \nTetra Ethylene Glycol 6.4 mls  \nCarboxylated Beads (5% solids.0.8um dia.) 2.0 mls  \nCommon Issues  \nTrapped Air Bubbles Can Prevent Mixing  \nEvaporation During Thermal Cycling Causes Low Read-Length Improper Mixing Causes Dropouts and Loss ofTrace Resolution  \nmagnetic particles with a solution of polyethylene glycol and sodium chloride. The beads were washed multiple times with 70% ethanol and pure DNA was eluted with water. While this method met the requirements listed above, we needed a technique that worked in 384-well PCR plates and produced extremely pure DNA.  \nOur first design consideration focused on replacing the polyethylene glycol and sodium chloride binding buffer. Sodium chloride inhibits electrokinetic injection and is significant source of variability in capi","cbCaiioJQoNTkkQq","https://ap.wps.com/l/cbCaiioJQoNTkkQq","pdf",662696,"English","# Abstract\n# Introduction\n# Material and Methods\n# Results and Discussion\n## Table 1. Sequencing Fragment Purification Protocol\n## Reagent/Stock Preparation\n## Common Issues\n# Reagent and Binding-Buffer Design Considerations","[{\"question\":\"What sequencing purification approach is described in the document?\",\"answer\":\"An automated magnetic bead purification workflow is used to clean terminator sequencing products in a 384-well format.\"},{\"question\":\"How does the magnetic bead method compare with ethanol precipitation?\",\"answer\":\"Compared with laborious ethanol precipitation, it increases phred20 read length by about forty bases while producing highly pure labeled DNA fragments.\"},{\"question\":\"Which capillary electrophoresis instruments are compatible with the method?\",\"answer\":\"The protocol is compatible with both the MegaBACE 1000 and ABI Prism 3700 capillary instruments.\"}]","Magnetic Bead Purification of Labeled DNA Fragments for High-Throughput Capillary Electrophoresis Sequencing | PDF",1789206422]