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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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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. |
format | Online Article Text |
id | pubmed-6080204 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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