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Monitoring of chicken RNA integrity as a function of prolonged postmortem duration
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Metadata
Document Title
Monitoring of chicken RNA integrity as a function of prolonged postmortem duration
Author
Malila Y., Srimarut Y., U-Chupaj J., Strasburg G., Visessanguan W.
Name from Authors Collection
Affiliations
National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Khlong Luang, Pathum Thani, 12120, Thailand; Department of Food Technology, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, Songkhla, 90112, Thailand; Department of Food Science and Human Nutrition, Michigan State University, East Lansing, MI 48824, United States
Type
Article
Source Title
Asian-Australasian Journal of Animal Sciences
ISSN
10112367
Year
2015
Volume
28
Issue
11
Page
1649-1656
Open Access
Gold, Green
Publisher
Asian-Australasian Association of Animal Production Societies
DOI
10.5713/ajas.15.0167
Abstract
Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect A260/A280 and A260/A230 (p.0.05), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples (p.0.05), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthineguanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meatquality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing. Copyright © 2015 by Asian-Australasian Journal of Animal Sciences.
Keyword
Gallus gallus | Gene Expression | Postmortem Duration | RNA Integrity | Skeletal Muscle
License
CC BY
Rights
Asian-Australasian Association of Animal Production Societies
Publication Source
Scopus