Table of Contents
Molecular Biology International
Volume 2013, Article ID 587680, 7 pages
Research Article

GAPDH Pseudogenes and the Quantification of Feline Genomic DNA Equivalents

Clinical Laboratory, Vetsuisse Faculty, University of Zurich, Winterthurerstrasse 260, 8057 Zurich, Switzerland

Received 17 January 2013; Revised 21 March 2013; Accepted 28 March 2013

Academic Editor: Emanuel Strehler

Copyright © 2013 A. Katrin Helfer-Hungerbuehler et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Linearity of the qPCR assay and absolute quantification of fALB amplification: The linearity of the qPCR assay was determined using a ten-fold serial dilution of the linearized standard template. In this Figure, representative results from a qPCR run using a Rotor-Gene 6000 real-time rotary analyzer (Corbett) are shown. A) Amplification plot of the fALB qPCR assay depicts cycle number versus normalized fluorescence. B) A standard curve (of a representative qPCR) shows the logarithmic starting input quantity (copies per reaction) of a 10-fold serial dilution of the standard template versus the measured cycle threshold (CT). The CT refers to the number of cycles required before the fluorescence passes a fixed threshold. Earlier increases in normalized fluorescence are associated with lower threshold cycle numbers and therefore higher starting quantities of sample template. The fALB assay showed linearity over 8 orders of magnitude. The correlation coefficient of the curve was 0.999, and the slope of the dilution versus threshold cycle curve was -3.33, which is ideal.

  1. Supplementary Material