expfit {qpcR} | R Documentation |
Calculation of PCR efficiency by fitting an exponential model
Description
An exponential model is fit to a window of defined size on the qPCR raw data. The window is identified either by the second derivative maximum 'cpD2' (default), 'studentized outlier' method as described in Tichopad et al. (2003), the 'midpoint' method (Peirson et al., 2003) or by subtracting the difference of cpD1 and cpD2 from cpD2 ('ERBCP', unpublished).
Usage
expfit(object, method = c("cpD2", "outlier", "midpoint", "ERBCP"),
model = c("exp", "linexp"), offset = 0, pval = 0.05, n.outl = 3,
n.ground = 1:5, corfact = 1, fix = c("top", "bottom", "middle"),
nfit = 5, plot = TRUE, ...)
Arguments
object |
an object of class 'pcrfit'. |
method |
one of the four possible methods to be used for defining the position of the fitting window. |
model |
which exponential model to use. |
offset |
for |
pval |
for |
n.outl |
for |
n.ground |
for |
corfact |
for |
fix |
for methods "midpoint" and "ERBCP", the orientation of the fitting window based on the identified point. See 'Details'. |
nfit |
the size of the fitting window. |
plot |
logical. If |
... |
other parameters to be passed to the plotting function. |
Details
The exponential growth function f(x) = a \cdot exp(b \cdot x) + c
is fit to a subset of the data. Calls efficiency
for calculation of the second derivative maximum, takeoff
for calculation of the studentized residuals and 'outlier' cycle, and midpoint
for calculation of the exponential phase 'midpoint'. For method 'ERBCP' (Exponential Region By Crossing Points), the exponential region is calculated by expR = cpD2 - \code{corfact} \cdot (cpD1-cpD2)
. The efficiency is calculated from the exponential fit with E = exp(b)
and the inital template fluorescence F_0 = a
.
Value
A list with the following components:
point |
the point within the exponential region as identified by one of the three methods. |
cycles |
the cycles of the identified region. |
eff |
the efficiency calculated from the exponential fit. |
AIC |
the Akaike Information Criterion of the fit. |
resVar |
the residual variance of the fit. |
RMSE |
the root-mean-squared-error of the fit. |
init |
the initial template fluorescence. |
mod |
the exponential model of class 'nls'. |
Author(s)
Andrej-Nikolai Spiess
References
Standardized determination of real-time PCR efficiency from a single reaction set-up.
Tichopad A, Dilger M, Schwarz G & Pfaffl MW.
Nucleic Acids Research (2003), 31:e122.
Comprehensive algorithm for quantitative real-time polymerase chain reaction.
Zhao S & Fernald RD.
J Comput Biol (2005), 12:1047-64.
Examples
## Using default SDM method.
m1 <- pcrfit(reps, 1, 2, l5)
expfit(m1)
## Using 'outlier' method.
expfit(m1, method = "outlier")
## Linear exponential model.
expfit(m1, model = "linexp")