Livestock Studies
2018, Vol 58, Num, 1 (Pages: 042-047)
RNA Quality in Snap and Directly -80°C Frozen Liver Tissues
Hüseyin Özkan 1 ,Akın Yakan 1
1 Mustafa Kemal Üniversitesi Veteriner Fakültesi Genetik AD Antakya/ Hatay
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Cells response can be analyzed at the level of mRNA with gene expression studies to physiological and biological changes. The stability of RNA, a highly sensitive molecule, varies from tissue to tissue. The RNA in the tissues can be rapidly degraded by exposure to nucleases after collecting from the species of animate. In order to control the degradation, the tissue should be frozen or treated with some chemicals. Nucleases are protein nature and they are inacitve at low temperatures so in many studies freezing method at directly -80°C with some chemical solutions or snap frozen with using liquid nitrogen method are preferred. This study was conducted to evaluate the concentration, purity and integrity criteria of nucleases from stock of liver tissues using different methods in rats. 8 rats which were six months old rats used in the study. The RNA isolated from liver tissues for Directly -80°C and Snap Frozen groups and examined of concentrations (1039,64 and 503,14 ng/μl), purity (1,98 and 1,97) and Ct values (16,537 and 17,463) of RNA in PPIA as reference gene. Consequently, RNA used in gene expression studies should been evaluated as a whole in terms of concentration, agarose gel image, melting curve and Ct values for quality assessment and the Snap Frozen method has been still considered one of the most appropriate method.
Keywords :
Snap Frozen, Directly -80°C, RNA quality, Gene expression