| Abstrakt: |
Contents The aims of this study were to test (i) the effect of time of tissue and RNA extracts storage on ice and (ii) the effect of repeated freeze-thaw cycles on RNA integrity and gene expression of bovine reproductive tissues. Fragments of endometrium ( ENDO), corpus luteum ( CL) and ampulla ( AMP) were subdivided and incubated for 0, 1, 3, 6, 12 or 24 h on ice. RNA extracts were incubated on ice for 0, 3, 12 or 24 h, or exposed to 1, 2, 4 or 6 freeze-thaw cycles. RNA integrity number ( RIN) was estimated. Expression of progesterone receptor ( PGR) and cyclophilin genes from RNA extracts stored on ice for 0 or 24 h, and 1 or 6 freeze-thaw cycles was measured by q PCR. Tissue and RNA extract incubation on ice, and repeated freeze-thaw cycles did not affect RIN values of RNA from ENDO, CL or AMP. Storage on ice or exposure to freeze-thaw cycles did not affect Cq values for PGR or cyclophilin genes . In conclusion, neither generalized RNA degradation nor specific RNA degradation was affected by storage of tissue or RNA extracts on ice for up to 24 h, or by up to 6 freeze-thaw cycles of RNA extracts obtained from bovine ENDO, CL and AMP. [ABSTRACT FROM AUTHOR] |
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