Type

Journal Article

Authors

Eamonn Gormley
Eamon Costello
David E. MacHugh
John A Browne
Michael P Sheridan

Subjects

Microbiology

Topics
time factors metabolism isolation purification animals interferon gamma reverse transcriptase polymerase chain reaction cattle genetics veterinary dna complementary gene expression profiling blood male rna messenger

Impact of delayed processing of bovine peripheral blood on differential gene expression. (2011)

Abstract RT-qPCR can be used to accurately determine expression levels of genes following RNA extraction from tissue samples. If blood is the source of total RNA, it is often desirable to process the samples immediately following collection because delays in processing for RNA extraction may influence mRNA expression estimates obtained from RT-qPCR analyses. However, this may not be feasible if the site of blood collection is distant from the processing laboratory. In the present study, the effects of delays in the processing of blood samples on mRNA expression data was investigated using a panel of 23 functionally diverse genes from five different gene ontology (GO) categories in peripheral blood sampled from ten age-matched healthy cattle. Venous blood was collected in Tempus™ Blood RNA tubes, which contain reagents that lyse blood cells immediately and stabilise the RNA signature (T(0)). Blood was also collected in conventional lithium heparin collection tubes, and stored at ambient temperature for T(4), T(6) and T(8)h, prior to total RNA extraction. The mRNA expression profiles of these 23 genes were determined by RT-qPCR and compared across the time course. Thirteen genes showed significant up- or down-fold changes in mRNA expression over the 8h time course. Among the GO categories, genes in the Immune response category showed the most differential expression. These results also demonstrated that the changes in mRNA expression for the IFNG gene, which encodes the cytokine IFN-γ, did not correspond to IFN-γ protein levels estimated using ELISA.
Collections Ireland -> University College Dublin -> PubMed

Full list of authors on original publication

Eamonn Gormley, Eamon Costello, David E. MacHugh, John A Browne, Michael P Sheridan

Experts in our system

1
E Gormley
University College Dublin
Total Publications: 68
 
2
Eamon Costello
Dublin City University
Total Publications: 94
 
3
David E. MacHugh
University College Dublin
Total Publications: 82
 
4
John A Browne
University College Dublin
Total Publications: 73