Abstract
RNAlater® is regarded as a potential preservation method for proteins, while its effect on bovine muscle proteins has rarely been evaluated. Bovine muscle protein samples (n = 12) collected from three tender (Warner–Bratzler shear force: 30.02–31.74 N) and three tough (Warner–Bratzler shear force: 54.12–66.25 N) Longissimus thoracis et lumborum (LTL) samples, preserved using two different sampling preservation methods (RNAlater® and dry ice), at two post mortem time points (day 0 and day 14), were characterized using one-dimensional electrophoresis. Fourteen bands with molecular weights ranging from 15 to 250 kDa were verified, both in the dry ice and RNAlater® storage groups, at each time point, using image analysis. A shift from high to low molecular weight fragments, between day 0 and day 14, indicated proteolysis of the muscle proteins during post mortem storage. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses and database searching resulted in the identification of 10 proteins in four bands. Protein profiles of muscle preserved in RNAlater® were similar to those of muscle frozen on dry ice storage, both at day 0 and day 14. The results demonstrate that RNAlater® could be a simple and efficient way to preserve bovine muscle proteins for bovine muscle proteomic studies.
Highlights
Meat proteomics research critically depends on the reliability of tissue samples
The objective of this study was to compare proteome patterns of post mortem bovine muscle stored in RNAlater® or dry ice by using one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (1D SDS-PAGE)
Fourteen bands were detected in each lane for both RNAlater® -preserved and dry ice-preserved samples
Summary
Meat proteomics research critically depends on the reliability of tissue samples. It is essential to avoid proteolysis and enzymatic activity in sample preservation, so as to conserve the structural integrity of muscle proteins. Snap freezing of samples in liquid nitrogen, at −196 ◦ C, or on dry ice, at −79 ◦ C, is an efficient method to stabilize samples. They can be difficult to carry out in-factory. RNAlater® , an aqueous solution, is extensively used to preserve RNA in fresh tissue and cell samples for clinical genomic and transcriptomic studies [1].
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