HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Pena, R. N. Articles by Estany, J. PubMed PubMed Citation Articles by Pena, R. N. Articles by Estany, J.

Technical Note: Efficient protocol for isolation of total RNA from lyophilised fat and muscle pig samples

R. N. Pena^*, A. Cánovas^* and J. Estany

^* IRTA, Genètica i Millora Animal, 191 Av Rovira Roure, 25198 Lleida, Spain Departament de Producció Animal, Universitat de Lleida, 191 Av Rovira Roure, 25198 Lleida, Spain

romi.pena@irta.cat

Abstract

Isolation of total RNA from frozen muscle and fat samples typically results in low yields due to the presence of connective tissue between muscle fibres, which impairs complete tissue homogenisation, and the excess of fat and low cellularity of adipose tissue. Meat quality studies involve determination of fatty acid composition and content from muscle and subcutaneous fat samples, a process which may produce an excess of lyophilised tissue samples. The purpose of this work was to investigate the stability of total RNA in lyophilised tissue samples generated during the routine detection of fatty acid content of pig muscle and fat tissues, stored either at room temperature or at -20ºC. The protocol described here results in high yields of total RNA from freeze-dried samples stored at -20ºC, which facilitates the homogenisation step. The isolated RNA is suitable for common gene expression techniques such as final point and quantitative reverse transcription-PCR.

Key Words: gene expression • freeze-dry • lyophilisation • pig • meat quality