Recovery of RNA from avian nucleated whole blood stored under field conditions: an evaluation of commercial methods. 2022

C Komini, and A Bounas
Department of Biological Applications and Technology, University of Ioannina, Ioannina, Greece.

1. Whole blood can be used to gain insights on several molecular mechanisms involved in animal physiology. However, its use to extract RNA from organisms that carry nucleated red blood cells is limited due to its high nuclease content and instability under field conditions. 2. Here, we evaluate three commercially available RNA preservation and extraction methods applied to avian whole-blood samples kept in different storage conditions to identify the most suitable ones for use in field studies. 3. Whole-blood starting volume was the most important factor; only samples with a starting volume of 50 µl yielded RNA, whereas all 100 µl samples failed to provide any RNA regardless of the method used. RNA concentration decreased with storage time in every method employed. Samples stored at -20°C yielded higher RNA concentration than the ones stored at 4°C, although some possible effects of freeze-thaw cycles were observed. 4. For two of the three methods, it is likely that DNA contamination and/or extensive degradation of RNA could have taken place. Only whole blood stored and extracted using the Quick RNA Whole Blood kit provided high-quality RNA in every condition tested.

UI MeSH Term Description Entries
D002645 Chickens Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA. Gallus gallus,Gallus domesticus,Gallus gallus domesticus,Chicken
D005615 Freezing Liquids transforming into solids by the removal of heat. Melting
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012313 RNA A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed) RNA, Non-Polyadenylated,Ribonucleic Acid,Gene Products, RNA,Non-Polyadenylated RNA,Acid, Ribonucleic,Non Polyadenylated RNA,RNA Gene Products,RNA, Non Polyadenylated

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