RNA Extraction from Liver Samples

RNA was extracted, DNaseI-treated, and purified from flash-frozen 50–100 mg liver samples following the standard procedures in the laboratory (Xu et al. 2013 (link); Xue et al. 2015 (link); Caballero-Solares et al. 2018 (link)). RNA integrity and purity were verified by 1% agarose gel electrophoresis and NanoDrop UV spectrophotometry (Thermo Fisher Scientific, Mississauga, ON, Canada), respectively. For all samples, no signs of RNA degradation were found (i.e., intact 28S and 18S ribosomal RNA bands), and A260/280 and A260/230 ratios were 2.1–2.3 and 1.9–2.4, respectively.

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Caballero-Solares A., Xue X., Cleveland B.M., Foroutani M.B., Parrish C.C., Taylor R.G, & Rise M.L. (2020). Diet-Induced Physiological Responses in the Liver of Atlantic Salmon (Salmo salar) Inferred Using Multiplex PCR Platforms. Marine Biotechnology (New York, N.y.), 22(4), 511-525.

Publication 2020

NanoDrop UV spectrophotometry

18s ribosomal rna Agarose gel electrophoresis Frozen Liver Rna degradation Spectrophotometry

Corresponding Organization : Memorial University of Newfoundland

Other organizations : National Center for Cool and Cold Water Aquaculture, Cargill (United States)

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Not explicitly mentioned

control variables

Flash-frozen 50–100 mg liver samples
RNA integrity and purity verified by 1% agarose gel electrophoresis and NanoDrop UV spectrophotometry
No signs of RNA degradation (intact 28S and 18S ribosomal RNA bands)
A260/280 and A260/230 ratios within acceptable ranges (2.1–2.3 and 1.9–2.4, respectively)

controls

Positive control: Not mentioned
Negative control: Not mentioned

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