C126EG685.Rd

\name{C126EG685}
\alias{C126EG685}
\docType{data}
\title{
qPCR Experiment for the Amplification of HPRT1 Using the Roche Bio-Rad iQ5
}
\description{
A Quantitive PCR (qPCR) with the DNA binding dye (EvaGreen) (Mao et al. 2007) 
was performed in the iQ5 (Bio-Rad) thermo cycler. The cycle-dependent increase 
of the fluorescence was quantified at the elongation step (68.5 deg Celsius).
}
\usage{data(C126EG685)}	
\format{
  A data frame with 40 observations on the following 97 variables. The first 
  column ("Cycles") contains the number of cylces and consecutive columns 
  contain the replicates ("A01" to "H12").
}
\details{
MLC-2v was amplified in the Roche Ligth Cycler 1.5. The the change of 
fluorescence was simultaniously monitored for the Hydrolysis probe of MLC-2v 
and EvaGreen. The primer sequences for MLC-2v were taken from Roediger et al. 
(2013). A 10 micro L qPCR reaction was composed of 250 nM primer (forward and 
reverse), qPCR Mix (accordingt to the manufactures recommendations), 1 micro L 
template  (MLC-2v amplification product), 60 nM hydorlysis probe probe for 
MLC-2v. EvaGreen was used at 0.5x final. The amplification was monitored at 
68.5 degree Celsius (elongation step).
}
\source{
Claudia Deutschmann & Stefan Roediger, BTU Cottbus - Senftenberg, Senftenberg, 
Germany
}
\references{
A Highly Versatile Microscope Imaging Technology Platform for the Multiplex 
Real-Time Detection of Biomolecules and Autoimmune Antibodies. S. Roediger, 
P. Schierack, A. Boehm, J. Nitschke, I. Berger, U. Froemmel, C. Schmidt, 
M. Ruhland, I. Schimke, D. Roggenbuck, W. Lehmann and C. Schroeder.
\emph{Advances in Biochemical Bioengineering/Biotechnology}. 133:33--74, 2013.
\url{http://www.ncbi.nlm.nih.gov/pubmed/22437246}

Mao, F., Leung, W.-Y., Xin, X., 2007. Characterization of EvaGreen and the 
implication of its physicochemical properties for qPCR applications. 
\emph{BMC Biotechnol}. 7, 76.
}
\examples{
data(C126EG685)
tmp <- C126EG685

plot(NA,NA, xlim = c(1,40), ylim = c(min(tmp[, 2:ncol(tmp)]), 
    max(tmp[, 2:ncol(tmp)])), xlab = "Cycle", 
    ylab = "RFU (FAM)", 
    main = "Amplification monitored at \n68.5 degree Celsius (elongation step)")

apply(tmp[, 2:ncol(tmp)], 2, 
      function(x) lines(tmp[1:nrow(tmp),1],x))
      
}
\keyword{ datasets }
\keyword{ EvaGreen }
\keyword{ HPRT1 }