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BioTechniques
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Extraction of ultrashort DNA molecules from herbarium specimens

Authors: Gutaker, Rafal M.; Reiter, Ella; Furtwängler, Anja; Schuenemann, Verena J.; Burbano, Hernán A.;

Extraction of ultrashort DNA molecules from herbarium specimens

Abstract

Abstract DNA extracted from herbarium specimens is highly fragmented and decays at a faster rate than DNA from ancient bones. Therefore, it is crucial to utilize extraction protocols that retrieve short DNA molecules. Improvements in extraction and library preparation protocols for animal remains have allowed efficient retrieval of molecules shorter than 50 bp. We adapted those improvements to extraction protocols for herbarium specimens and evaluated their performance by shotgun sequencing, which allows an accurate estimation of the distribution of fragment lengths. Extraction with PTB buffer decreased median fragment length by 35% when compared to CTAB. Modifying the binding conditions of DNA to silica allowed for an additional decrease of 10%. We did not observe a further decrease in length when we used single-stranded instead of double-stranded library preparation methods. Our protocol enables the retrieval of ultrashort molecules from herbarium specimens and will help to unlock the genetic information stored in herbaria.

Keywords

DNA, Plant, QH301-705.5, Cetrimonium, Sequence Analysis, DNA, Polymerase Chain Reaction, Thiazoles, paleogenomics, Cetrimonium Compounds, next-generation sequencing, Biology (General), DNA, Ancient, ancient DNA, DNA extraction, DNA extraction ; herbarium ; next-generation sequencing ; paleogenomics ; ancient DNA, herbarium, Gene Library

  • BIP!
    Impact byBIP!
    selected citations
    These citations are derived from selected sources.
    This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    51
    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Top 10%
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    Top 10%
    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Top 10%
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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
51
Top 10%
Top 10%
Top 10%
Green
gold