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DNA Extraction, Preservation, and Amplification

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Book cover DNA Barcodes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 858))

Abstract

The effectiveness of DNA barcoding as a routine practice in biodiversity research is strongly dependent on the quality of the source material, DNA extraction method, and selection of adequate primers in combination with optimized polymerase chain reaction (PCR) conditions. For the isolation of nucleic acids, silica-gel membrane methods are to be favored because they are easy to handle, applicable for high sample throughput, relatively inexpensive, and provide high DNA quality, quantity, and purity which are pre­requisites for successful PCR amplification and long-term storage of nucleic acids in biorepositories, such as DNA banks. In this section, standard protocols and workflow schemes for sample preparation, DNA isolation, DNA storage, PCR amplification, PCR product quality control, and PCR product cleanup are proposed and described in detail. A PCR troubleshooting and primer design section may help to solve problems that hinder successful amplification of the desired barcoding gene region.

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Acknowledgment

This work was supported by the German Science Foundation DFG as part of the DNA Bank Network project (http://www.dnabank-network.org).

Author information

Authors and Affiliations

  1. German Centre for Marine Biodiversity Research (DZMB), Senckenberg Research Institute, Wilhelmshaven, Germany

    Thomas Knebelsberger

  2. Bavarian State Collection of Zoology (ZSM), Munich, Germany

    Isabella Stöger

Authors
  1. Thomas Knebelsberger
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  2. Isabella Stöger
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Corresponding author

Correspondence to Thomas Knebelsberger .

Editor information

Editors and Affiliations

  1. National Museum of Natural History, Dept. Botany, Smithsonian Institution, 10th St. & Constitution Ave. NW.,, Washington, 20560-0166, District of Columbia, USA

    W. John Kress

  2. National Museum of Natural History, Dept. Botany, Smithsonian Institution, 10th St. & Constitution Ave. NW.,, Washington, 20560-0166, District of Columbia, USA

    David L. Erickson

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Knebelsberger, T., Stöger, I. (2012). DNA Extraction, Preservation, and Amplification. In: Kress, W., Erickson, D. (eds) DNA Barcodes. Methods in Molecular Biology, vol 858. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-591-6_14

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  • DOI: https://doi.org/10.1007/978-1-61779-591-6_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-590-9

  • Online ISBN: 978-1-61779-591-6

  • eBook Packages: Springer Protocols

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