Semi-automated library preparation for high-throughput DNA sequencing platforms.
Next-generation sequencing platforms are powerful technologies, providing gigabases of genetic information in a single run. An important prerequisite for high-throughput DNA sequencing is the development of robust and cost-effective preprocessing protocols for DNA sample library construction. Here w...
| Publicado en: | Journal of Biomedicine & Biotechnology pp. 8p - 9 |
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| Autores principales: | , , , , , , |
| Formato: | research tables/charts Journal Article |
| Publicado: |
Wiley-Blackwell
2010
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=105082287&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 105082287 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 11107243 137K jtl: Journal of Biomedicine & Biotechnology issn: 11107243 maglogo: N pubinfo: dt: 2010 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 105082287 2010748642 10.1155/2010/617469 NLM20625503 PMC2896710 105082287 ppf: 8p ppct: 1 formats: fmt: @attributes: type: P tig: atl: Semi-automated library preparation for high-throughput DNA sequencing platforms. aug: au: Farias-Hesson E Erikson J Atkins A Shen P Davis RW Scharfe C Pourmand N affil: Department of Biomolecular Engineering, University of California, Santa Cruz, CA 95064, USA. sug: subj: DNA Libraries Computers and Computerization Funding Source Genetic Techniques Methods Human Mitochondria Protocols Robotics ab: Next-generation sequencing platforms are powerful technologies, providing gigabases of genetic information in a single run. An important prerequisite for high-throughput DNA sequencing is the development of robust and cost-effective preprocessing protocols for DNA sample library construction. Here we report the development of a semi-automated sample preparation protocol to produce adaptor-ligated fragment libraries. Using a liquid-handling robot in conjunction with Carboxy Terminated Magnetic Beads, we labeled each library sample using a unique 6 bp DNA barcode, which allowed multiplex sample processing and sequencing of 32 libraries in a single run using Applied Biosystems' SOLiD sequencer. We applied our semi-automated pipeline to targeted medical resequencing of nuclear candidate genes in individuals affected by mitochondrial disorders. This novel method is capable of preparing as much as 32 DNA libraries in 2.01 days (8-hour workday) for emulsion PCR/high throughput DNA sequencing, increasing sample preparation production by 8-fold. pubtype: Academic Journal doctype: research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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