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

Abstract 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 is...

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Bibliographic Details
Published in:BMC Research Notes
Main Authors: Jason A. Ferrante, Michelle R. Giles, Emily Benzie, Margaret E. Hunter
Format: Article in Journal/Newspaper
Language:English
Published: BMC 2018
Subjects:
Blood
Tempus
RNA
Genomic DNA
Isolation
Preservation
Medicine
R
Biology (General)
QH301-705.5
Science (General)
Q1-390
Alces alces
Online Access:https://doi.org/10.1186/s13104-018-3671-4
https://doaj.org/article/430b62901db74bfda4b240caa085c541
Description
Summary:Abstract 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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