Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

A quantitative method for comparison of expression of alternatively spliced genes using different primer pairs

Donev, Rossen Mintchev and Morgan, Bryan Paul 2006. A quantitative method for comparison of expression of alternatively spliced genes using different primer pairs. Journal of Biochemical and Biophysical Methods 66 (1-3) , pp. 23-31. 10.1016/j.jbbm.2005.11.001

Full text not available from this repository.

Abstract

In this paper we describe a novel two-step method for comparison of expression of alternatively spliced genes by quantitative PCR (QPCR) applying different primer pairs. As a model system we used rat decay accelerating factor (DAF; CD55) mRNA, which comprises three different isoforms: soluble (sDAF), transmembrane (tmDAF) and glycosyl-phosphatidylinositol (GPI) anchored (gpiDAF) forms. The first step was to prepare solid phase specific for each mRNA isoform and purify the three DAF-forms from total RNA. We then assessed amplification efficiency of primer pairs designed to recognise each of the isoforms using equimolar amounts of the three purified DAF mRNAs. The final step in our assay was to compare expression of the three DAF-isoforms in testis by QPCR taking into account the efficiency of their amplification to enable quantification. The RNA capture/QPCR method we described here can be used for quantifying the expression ratios of alternatively spliced mRNAs from a single gene or for direct comparison of expression of different genes.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Subjects: R Medicine > R Medicine (General)
Publisher: Elsevier
ISSN: 0165-022X
Last Modified: 04 Jun 2017 06:41
URI: http://orca-mwe.cf.ac.uk/id/eprint/63636

Citation Data

Cited 5 times in Google Scholar. View in Google Scholar

Cited 4 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item Edit Item