1. A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation
- Author
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Michelle R. Giles, Margaret E. Hunter, Emily Benzie, and Jason A. Ferrante
- Subjects
0301 basic medicine ,Novel technique ,lcsh:Medicine ,Genomic DNA ,Wildlife ,General Biochemistry, Genetics and Molecular Biology ,Isolation ,03 medical and health sciences ,chemistry.chemical_compound ,Animals ,A-DNA ,lcsh:Science (General) ,lcsh:QH301-705.5 ,Chemistry ,Deer ,lcsh:R ,RNA ,General Medicine ,DNA ,Tempus ,Isolation (microbiology) ,Molecular biology ,Preservation ,genomic DNA ,Research Note ,030104 developmental biology ,Blood ,lcsh:Biology (General) ,Agarose ,RNA extraction ,lcsh:Q1-390 - 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.
- Published
- 2018