C54: qPCR Experiment for the amplification of MLC-2v using the...

In chipPCR: Toolkit of Helper Functions to Pre-Process Amplification Data

Description

qPCR Experiment for the amplification of MLC-2v using the 'VideoScan' heating/cooling-unit

Usage

data(C54)

Format

A data frame with 56 observations on the following 4 variables.

Cycle

Cycle number

D1

Stock concentration of input cDNA

D2

1/10 diluted stock cDNA

D3

1/100 diluted stock cDNA

Details

The aim was to amplify MLC-2v in the 'VideoScan' and to monitor with a hydrolysis probe for MLC-2v. The primer sequences for MLC-2v were taken from Roediger et al. (2013). The amplification was detected in solution of the '1 HCU' (see Roediger et al. 2013 for details). A 20 micro L PCR reaction was composed of 250 nM primer (forward and reverse), 1x Maxima Probe qPCR Master Mix (Fermentas), 1 micro L template (MLC-2v amplification product in different dilutions), 50 nM hydrolysis probe probe for MLC-2v and A. bidest. During the amplification, fluorescence was measured at 59.5 degree Celsius. The Cy5 channel was used to monitor the MLC-2v specific hydrolysis probe. Input stock cDNA was used undiluted (D1). D2 was 1/1000 and D3 1/1000000 diluted in A. bidest. The D1, D2, and D3 have different numbers measure points and D2 contains a missing value at cycle 37.

Source

Stefan Roediger, Claudia Deutschmann

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. Advances in Biochemical Bioengineering/Biotechnology. 133:33–74, 2013.

Examples

data(C54)
str(C54)
plot(NA, NA, xlim = c(0,56), ylim = c(0, 0.7), xlab = "Cycle", 
     ylab = "refMFI")
apply(C54[, c(2:4)], 2, function(x) lines(C54[, 1], x))

chipPCR documentation built on March 5, 2021, 9:06 a.m.