Storage of Bovine Reproductive Tissues and RNA Extracts on Ice for 24 h or Repeated Freeze–Thaw Cycles do not Affect RNA Integrity

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 subd...

Full description

Saved in:
Bibliographic Details
Published in:Reproduction in domestic animals 2014-02, Vol.49 (1), p.e9-e11
Main Authors: França, MR, Mesquita, FS, Binelli, M
Format: Article
Language:English
Subjects:
Animal reproduction
Animals
Cattle
Cold Temperature
corpus luteum
Corpus Luteum - chemistry
Corpus Luteum - physiology
Cryopreservation - methods
Cryopreservation - veterinary
Cyclophilins - chemistry
Endometrium - chemistry
Endometrium - physiology
Fallopian Tubes - chemistry
Fallopian Tubes - physiology
Female
Gene Expression
genes
Ice
progesterone
quantitative polymerase chain reaction
Receptors, Progesterone - genetics
Ribonucleic acid
RNA
RNA - genetics
RNA - isolation & purification
Tissue Preservation - methods
Tissue Preservation - veterinary
Tissues
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 qPCR. 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.
ISSN:0936-6768
1439-0531
DOI:10.1111/rda.12257