Development of a 3’:5’ digital PCR assay to determine horse mRNA integrity
Publication Name
Analytical Biochemistry
Abstract
Accurate tools to measure RNA integrity are essential to obtain reliable gene expression data. The reverse transcription quantitative PCR (RT-qPCR) based 3’:5′ assay permits a direct determination of messenger RNA (mRNA) integrity. However, the use of standard curves and the possible effect of PCR inhibitors make this method cumbersome and prone to variation, especially in small samples. Here we developed a triplex digital PCR (dPCR) 3’:5’ assay for assessing RNA integrity in equine samples as rapid and simple alternative to RT-qPCR. This dPCR assay not only provides a straight forward analysis of the mRNA integrity, but also of its quantity.
Open Access Status
This publication is not available as open access
Volume
626
Article Number
114217
Funding Number
FWO-SB 1S22820N
Funding Sponsor
ASCRS Research Foundation