Cargando…

Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens

Here we present and justify an approach for minimal-destructive DNA extraction from historic insect specimens for next generation sequencing applications. An increasing number of studies use insects from museum collections for biodiversity research. However, the availability of specimens for molecul...

Descripción completa

Detalles Bibliográficos
Autores principales: Patzold, Franziska, Zilli, Alberto, Hundsdoerfer, Anna K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7343169/
https://www.ncbi.nlm.nih.gov/pubmed/32639972
http://dx.doi.org/10.1371/journal.pone.0235222
_version_ 1783555716329504768
author Patzold, Franziska
Zilli, Alberto
Hundsdoerfer, Anna K.
author_facet Patzold, Franziska
Zilli, Alberto
Hundsdoerfer, Anna K.
author_sort Patzold, Franziska
collection PubMed
description Here we present and justify an approach for minimal-destructive DNA extraction from historic insect specimens for next generation sequencing applications. An increasing number of studies use insects from museum collections for biodiversity research. However, the availability of specimens for molecular analyses has been limited by the degraded nature of the DNA gained from century-old museum material and the consumptive nature of most DNA extraction procedures. The method described in this manuscript enabled us to successfully extract DNA from specimens as old as 241 years using a minimal-destructive approach. The direct comparison of the DNeasy extraction Kit and the Monarch® PCR & DNA Clean-up Kit showed a significant increase of 17.3-fold higher DNA yield extracted with the Monarch Oligo protocol on average. By using an extraction protocol originally designed for oligonucleotide clean-up, we were able to combine overcoming the restrictions by target fragment size and strand state, with minimising time consumption and labour-intensity. The type specimens used for the minimal-destructive DNA extraction exhibited no significant external change or post-extraction damage, while sufficient DNA was retrieved for analyses.
format Online
Article
Text
id pubmed-7343169
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-73431692020-07-17 Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens Patzold, Franziska Zilli, Alberto Hundsdoerfer, Anna K. PLoS One Research Article Here we present and justify an approach for minimal-destructive DNA extraction from historic insect specimens for next generation sequencing applications. An increasing number of studies use insects from museum collections for biodiversity research. However, the availability of specimens for molecular analyses has been limited by the degraded nature of the DNA gained from century-old museum material and the consumptive nature of most DNA extraction procedures. The method described in this manuscript enabled us to successfully extract DNA from specimens as old as 241 years using a minimal-destructive approach. The direct comparison of the DNeasy extraction Kit and the Monarch® PCR & DNA Clean-up Kit showed a significant increase of 17.3-fold higher DNA yield extracted with the Monarch Oligo protocol on average. By using an extraction protocol originally designed for oligonucleotide clean-up, we were able to combine overcoming the restrictions by target fragment size and strand state, with minimising time consumption and labour-intensity. The type specimens used for the minimal-destructive DNA extraction exhibited no significant external change or post-extraction damage, while sufficient DNA was retrieved for analyses. Public Library of Science 2020-07-08 /pmc/articles/PMC7343169/ /pubmed/32639972 http://dx.doi.org/10.1371/journal.pone.0235222 Text en © 2020 Patzold et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Patzold, Franziska
Zilli, Alberto
Hundsdoerfer, Anna K.
Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens
title Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens
title_full Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens
title_fullStr Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens
title_full_unstemmed Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens
title_short Advantages of an easy-to-use DNA extraction method for minimal-destructive analysis of collection specimens
title_sort advantages of an easy-to-use dna extraction method for minimal-destructive analysis of collection specimens
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7343169/
https://www.ncbi.nlm.nih.gov/pubmed/32639972
http://dx.doi.org/10.1371/journal.pone.0235222
work_keys_str_mv AT patzoldfranziska advantagesofaneasytousednaextractionmethodforminimaldestructiveanalysisofcollectionspecimens
AT zillialberto advantagesofaneasytousednaextractionmethodforminimaldestructiveanalysisofcollectionspecimens
AT hundsdoerferannak advantagesofaneasytousednaextractionmethodforminimaldestructiveanalysisofcollectionspecimens