The objective of this study was to evaluate the effect of post mortem times on the quality of porcine skeletal muscle total RNA in order to consider the possibility to use post mortem material for gene expression analysis. Samples of Musculus semimembranosus were collected at 20 min, 2 h, 6 h, 24 h and 48 h post mortem from the left legs of four commercial heavy pigs. Total RNA was analysed by agarose gel electrophoresis stained with ethidium bromide and by microfluidic capillary electrophoresis on an Agilent 2100 Bioanalyzer instrument obtaining 28S:18S rRNA peak ratios and RIN values. The average RIN values of the analysed samples were 7.45 ± 0.13, 7.43 ± 0.15, 7.45 ± 0.10, 7.33 ± 0.15 and 3.95 ± 0.58 for the same post mortem times, respectively, indicating that RNA degradation was present at 48 h post mortem. In a similar experiment, carried out by other authors on beef cattle muscle total RNA extracted at different post mortem times, RNA was stable up to 8 days after death as indicated by intact 28S and 18S rRNA bands. Thus, differences among species or other environmental factors might affect the level of RNA degradation. In the porcine post mortem samples, qualitative assessment of GAPDH transcripts by PCR amplification of different cDNA fragments indicated that post mortem stages did not affect the possibility of analysing this housekeeping gene. Thus, post mortem porcine skeletal muscle can be an useful tissue to obtain gene expression based information.

Evaluation of post mortem stability of porcine skeletal muscle RNA.

FONTANESI, LUCA;COLOMBO, MICHELA;BERETTI, FRANCESCA;RUSSO, VINCENZO
2008

Abstract

The objective of this study was to evaluate the effect of post mortem times on the quality of porcine skeletal muscle total RNA in order to consider the possibility to use post mortem material for gene expression analysis. Samples of Musculus semimembranosus were collected at 20 min, 2 h, 6 h, 24 h and 48 h post mortem from the left legs of four commercial heavy pigs. Total RNA was analysed by agarose gel electrophoresis stained with ethidium bromide and by microfluidic capillary electrophoresis on an Agilent 2100 Bioanalyzer instrument obtaining 28S:18S rRNA peak ratios and RIN values. The average RIN values of the analysed samples were 7.45 ± 0.13, 7.43 ± 0.15, 7.45 ± 0.10, 7.33 ± 0.15 and 3.95 ± 0.58 for the same post mortem times, respectively, indicating that RNA degradation was present at 48 h post mortem. In a similar experiment, carried out by other authors on beef cattle muscle total RNA extracted at different post mortem times, RNA was stable up to 8 days after death as indicated by intact 28S and 18S rRNA bands. Thus, differences among species or other environmental factors might affect the level of RNA degradation. In the porcine post mortem samples, qualitative assessment of GAPDH transcripts by PCR amplification of different cDNA fragments indicated that post mortem stages did not affect the possibility of analysing this housekeeping gene. Thus, post mortem porcine skeletal muscle can be an useful tissue to obtain gene expression based information.
MEAT SCIENCE
Fontanesi L.; Colombo M.; Beretti F.; Russo V.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/63488
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