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| Verfasst von: | Paruzynski, Anna [VerfasserIn]  |
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| | Arens, Anne [VerfasserIn] |
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| | Gabriel, Richard [VerfasserIn] |
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| | Bartholomä, Cynthia C. [VerfasserIn] |
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| | Scholz, Simone [VerfasserIn] |
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| | Wang, Wei [VerfasserIn] |
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| | Wolf, Stephan [VerfasserIn] |
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| | Glimm, Hanno [VerfasserIn] |
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| | Schmidt, Manfred [VerfasserIn] |
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| | Kalle, Christof von [VerfasserIn] |
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| Titel: | Genome-wide high-throughput integrome analyses by nrLAM-PCR and next-generation sequencing |
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| Verf.angabe: | Anna Paruzynski, Anne Arens, Richard Gabriel, Cynthia C. Bartholomae, Simone Scholz, Wei Wang, Stephan Wolf, Hanno Glimm, Manfred Schmidt & Christof von Kalle |
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| E-Jahr: | 2010 |
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| Jahr: | 08 July 2010 |
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| Umfang: | 10 S. |
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| Fussnoten: | Gesehen am 26.06.2023 |
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| Titel Quelle: | Enthalten in: Nature protocols |
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| Ort Quelle: | Basingstoke : Nature Publishing Group, 2006 |
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| Jahr Quelle: | 2010 |
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| Band/Heft Quelle: | 5(2010), 8, Seite 1379-1395 |
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| ISSN Quelle: | 1750-2799 |
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| Abstract: | High-throughput integration site profiling has become a feasible tool to assess vector biosafety and to monitor the cell fate of the gene-corrected cell population in clinical gene therapy studies. Here we report a step-by-step protocol for universal genome-wide and comprehensive integrome analysis that can be performed on >102-103 samples of interest in parallel. This assay is composed of fast and cost-efficient non-restrictive linear amplification-mediated PCR; optimized sample preparation for pyrosequencing; and automated bioinformatic data mining, including sequence trimming, alignment to the cellular genome and further annotation. Moreover, the workflow of this large-scale assay can be adapted to any PCR-based method aiming to characterize unknown flanking DNA adjacent to a known DNA region. Thus, in combination with next-generation sequencing technologies, large-scale integrome analysis of >4 × 105-1 × 106 integration site sequences can be accomplished within a single week. |
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| DOI: | doi:10.1038/nprot.2010.87 |
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| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
Volltext: https://doi.org/10.1038/nprot.2010.87 |
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| | Volltext: https://www.nature.com/articles/nprot.2010.87 |
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| | DOI: https://doi.org/10.1038/nprot.2010.87 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | Bioinformatics |
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| | Next-generation sequencing |
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| | Whole genome amplification |
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| K10plus-PPN: | 1850975876 |
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| Verknüpfungen: | → Zeitschrift |
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Genome-wide high-throughput integrome analyses by nrLAM-PCR and next-generation sequencing / Paruzynski, Anna [VerfasserIn]; 08 July 2010 (Online-Ressource)
