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  Creation and application of immortalized bait libraries for targeted enrichment and next-generation sequencing

Querfurth, R., Fischer, A., Schweiger, M. R., Lehrach, H., & Mertes, F. (2012). Creation and application of immortalized bait libraries for targeted enrichment and next-generation sequencing. Biotechniques, 52(6), 375-380. doi:10.2144/0000113877.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-000E-F07E-2 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-000E-F07F-F
Genre: Journal Article

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© 2012 BioTechniques
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 Creators:
Querfurth, Robert1, Author              
Fischer, Axel2, Author              
Schweiger, Michal R.2, Author              
Lehrach, Hans3, Author              
Mertes, Florian3, Author              
Affiliations:
1Dept. of Computational Molecular Biology (Head: Martin Vingron), Max Planck Institute for Molecular Genetics, Max Planck Society, Berlin, Germany, ou_1433547              
2Cancer Genomics (Michal-Ruth Schweiger), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, Berlin, Germany, ou_1479649              
3Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, Berlin, Germany, ou_1433550              

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Free keywords: Chromosome Mapping/methods Computational Biology/*methods Databases, Genetic *Gene Library Genes, Neoplasm Genetic Loci Humans Oligonucleotides/genetics Polymerase Chain Reaction Polymorphism, Single Nucleotide Sequence Analysis, DNA/*methods
 Abstract: Since the introduction of next-generation sequencing, several techniques have been developed to selectively enrich and sequence specific parts of the genome at high coverage. These techniques include enzymatic methods employing molecular inversion probes, PCR based approaches, hybrid capture, and in-solution capture. In-solution capture employs RNA probes transcribed from a pool of DNA template oligos designed to match regions of interest to specifically bind and enrich genomic DNA fragments. This method is highly efficient, especially if genomic target regions are large in size or quantity. Diverse in-solution capture kits are available, but are costly when large sample numbers need to be analyzed. Here we present a cost-effective strategy for the design of custom DNA libraries, their transcription into RNA libraries, and application for in-solution capture. We show the efficacy by comparing the method to a commercial kit and further demonstrate that emulsion PCR can be used for bias free amplification and virtual immortalization of DNA template libraries.

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Language(s): eng - English
 Dates: 2012-06
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.2144/0000113877
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Title: Biotechniques
  Other : Biotechniques
Source Genre: Journal
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Publ. Info: Natick, MA : Eaton Pub. Co.
Pages: - Volume / Issue: 52 (6) Sequence Number: - Start / End Page: 375 - 380 Identifier: ISSN: 0736-6205
CoNE: https://pure.mpg.de/cone/journals/resource/954925536105