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A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation

OBJECTIVE: We use Tempus blood RNA tubes (Applied Biosystems) during health assessments of American moose (Alces alces spp.) as a minimally invasive means to obtain RNA. Here we describe a novel protocol to additionally isolate high-quality DNA from the supernatant remaining after the RNA isolation...

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Detalles Bibliográficos
Autores principales: Ferrante, Jason A., Giles, Michelle R., Benzie, Emily, Hunter, Margaret E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6080204/
https://www.ncbi.nlm.nih.gov/pubmed/30081941
http://dx.doi.org/10.1186/s13104-018-3671-4
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author Ferrante, Jason A.
Giles, Michelle R.
Benzie, Emily
Hunter, Margaret E.
author_facet Ferrante, Jason A.
Giles, Michelle R.
Benzie, Emily
Hunter, Margaret E.
author_sort Ferrante, Jason A.
collection PubMed
description OBJECTIVE: We use Tempus blood RNA tubes (Applied Biosystems) during health assessments of American moose (Alces alces spp.) as a minimally invasive means to obtain RNA. Here we describe a novel protocol to additionally isolate high-quality DNA from the supernatant remaining after the RNA isolation methodology. Metrics used to qualify DNA quality included measuring the concentration, obtaining a DNA integrity number from a genomic DNA ScreenTape assay (Agilent), and running the isolated DNA on an agarose gel. RESULTS: Of the 23 samples analyzed, the average DNA concentration was 121 ng/µl (range 4–337 ng/µl) and a genomic DNA ScreenTape assay of seven samples indicated high DNA integrity values for 6 of the 7 samples (range 9.1–9.4 out of 10). Of the DNA sent for genotyping by sequencing, all proved to be of sufficient integrity to yield high-quality next-generation sequence results. We recommend this simple procedure to maximize the yield of both RNA and DNA from blood samples.
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spelling pubmed-60802042018-08-09 A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation Ferrante, Jason A. Giles, Michelle R. Benzie, Emily Hunter, Margaret E. BMC Res Notes Research Note OBJECTIVE: We use Tempus blood RNA tubes (Applied Biosystems) during health assessments of American moose (Alces alces spp.) as a minimally invasive means to obtain RNA. Here we describe a novel protocol to additionally isolate high-quality DNA from the supernatant remaining after the RNA isolation methodology. Metrics used to qualify DNA quality included measuring the concentration, obtaining a DNA integrity number from a genomic DNA ScreenTape assay (Agilent), and running the isolated DNA on an agarose gel. RESULTS: Of the 23 samples analyzed, the average DNA concentration was 121 ng/µl (range 4–337 ng/µl) and a genomic DNA ScreenTape assay of seven samples indicated high DNA integrity values for 6 of the 7 samples (range 9.1–9.4 out of 10). Of the DNA sent for genotyping by sequencing, all proved to be of sufficient integrity to yield high-quality next-generation sequence results. We recommend this simple procedure to maximize the yield of both RNA and DNA from blood samples. BioMed Central 2018-08-06 /pmc/articles/PMC6080204/ /pubmed/30081941 http://dx.doi.org/10.1186/s13104-018-3671-4 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Note
Ferrante, Jason A.
Giles, Michelle R.
Benzie, Emily
Hunter, Margaret E.
A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation
title A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation
title_full A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation
title_fullStr A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation
title_full_unstemmed A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation
title_short A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation
title_sort novel technique for isolating dna from tempus™ blood rna tubes after rna isolation
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6080204/
https://www.ncbi.nlm.nih.gov/pubmed/30081941
http://dx.doi.org/10.1186/s13104-018-3671-4
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