library(qtl). You may wish to include this in a
.Rprofile file. help.start function to start the
html version of the R help. library(help=qtl) to get a list of the functions
in R/qtl. example function to run examples
of the various functions in R/qtl. read.cross. The
internal data structure used by R/qtl is rather complicated, and is
described in the help file for read.cross. We won't
discuss data import any further here, except to say that the
comma-delimited format ("csv") is recommended. If you have
trouble importing data, send an email to Karl Broman,
kbroman@biostat.wisc.edu, perhaps attaching examples of your data
files. (Such data will be kept confidential.) Also see the sample data
files and code at http://www.rqtl.org/sampledata. We consider the example data hyper, an experiment on
hypertension in the mouse, kindly provided
by Bev Paigen and Gary Churchill. Use the data
function to load the data. data(hyper) The hyper data set has class "cross". The
function summary.cross gives summary information
on the data, and checks the data for internal consistency. A number
of other utility functions are available; hopefully these are
self-explanatory. summary(hyper)
nind(hyper)
nphe(hyper)
nchr(hyper)
nmar(hyper)
totmar(hyper) The function plot.cross gives a graphical summary of
the data; it calls plotMissing (to plot a matrix
displaying missing genotypes) and plotMap (to plot
the genetic maps), and also displays histograms or barplots of the
phenotypes. The plotMissing function can plot
individuals ordered by their phenotypes; you can see that for most
markers, only individuals with extreme phenotypes were genotyped. plot(hyper)
plotMissing(hyper)
plotMissing(hyper, reorder=TRUE)
plotMap(hyper) Note that one marker (on chromosome 14) has no genotype data. The
function drop.nullmarkers removes such markers from
the data. hyper <- drop.nullmarkers(hyper)
totmar(hyper) The function est.rf estimates the recombination
fraction between each pair of markers, and calculates a LOD score for
the test of $r$ = 1/2. This is useful for identifying markers that
are placed on the wrong chromosome. Note that since, for these data,
many markers were typed only on recombinant individuals, the pairwise
recombination fractions show rather odd patterns. hyper <- est.rf(hyper)
plotRF(hyper)
plotRF(hyper, chr=c(1,4)) To re-estimate the genetic map for an experimental cross, use the
function est.map. The function
plotMap, in addition to plotting a single map, can
plot the comparison of two genetic maps (as long as they are composed of
the same numbers of chromosomes and markers per chromosome). The
function replace.map map be used to replace the
genetic map in a cross with a new one. newmap <- est.map(hyper, error.prob=0.01, verbose=TRUE)
plotMap(hyper, newmap)
hyper <- replace.map(hyper, newmap) The function calc.errorlod may be used to assist in
identifying possible genotyping errors; it calculates the error LOD
scores described by Lincoln and Lander (1992). The
calc.errorlod function return a modified version of
the input cross, with error LOD scores included. The function
top.errorlod prints the genotypes with values above a
cutoff (by default, the cutoff is 4.0). hyper <- calc.errorlod(hyper, error.prob=0.01)
top.errorlod(hyper) The function plotGeno may be used to inspect the
observed genotypes for a chromosome, with likely genotyping errors
flagged. plotGeno(hyper, chr=16, ind=c(24:34, 71:81)) Before doing QTL analyses, some intermediate calculations need to be
performed. The function calc.genoprob calculates
conditional genotype probabilities given the multipoint marker data.
sim.geno simulates sequences of genotypes from their
joint distribution, given the observed marker data. As with calc.errorlod, these functions return a
modified version of the input cross, with the intermediate calculations
included. The step argument indicates the density of the grid on
which the calculations will be performed, and determines the density at
which LOD scores will be calculated. hyper <- calc.genoprob(hyper, step=2.5, error.prob=0.01)
hyper <- sim.geno(hyper, step=2.5, n.draws=64, error.prob=0.01) The function scanone performs a genome scan with a
single QTL model. By default, it performs standard interval mapping
(Lander and Botstein 1989): use of a normal model and the EM algorithm.
If one specifies method="hk", Haley-Knott regression is performed
(Haley and Knott 1992). These two methods require the results from
calc.genoprob. out.em <- scanone(hyper)
out.hk <- scanone(hyper, method="hk") If one specifies method="imp", a genome scan is performed by the
multiple imputation method of Sen and Churchill (2001). This method
requires the results from sim.geno. out.imp <- scanone(hyper, method="imp") The output of scanone is a data.frame with class
"scanone". The function plot.scanone may be
used to plot the results, and may plot up to three sets of results
against each other, as long as they conform appropriately. plot(out.em)
plot(out.hk, col="blue", add=TRUE)
plot(out.imp, col="red", add=TRUE)
plot(out.hk, out.imp, out.em, chr=c(1,4), lty=1,
col=c("blue","red","black")) The function summary.scanone may be used to list
information on the peak LOD for each chromosome for which the LOD
exceeds a specified threshold. summary(out.em)
summary(out.em, threshold=3)
summary(out.hk, threshold=3)
summary(out.imp, threshold=3) The function max.scanone returns the maximum LOD
score, genome-wide. max(out.em)
max(out.hk)
max(out.imp) One may also use scanone to perform a permutation
test to get a genome-wide LOD significance threshold. operm.hk <- scanone(hyper, method="hk", n.perm=1000) The result has class "scanoneperm". The
summary.scanoneperm function may be used to calculate
LOD thresholds. summary(operm.hk, alpha=0.05) The permutation results may also be used in the
summary.scanone function to calculate LOD thresholds
and genome-scan-adjusted p-values. summary(out.hk, perms=operm.hk, alpha=0.05, pvalues=TRUE) We should say at this point that the function
save.image will save your workspace to disk. You'll
wish you had used this if R crashes. save.image() The function scantwo performs a two-dimensional
genome scan with a two-QTL model. Methods "em", "hk" and
"imp" are all available. scantwo is
considerably slower than scanone, and can require a
great deal of memory. Thus, you may wish to re-run
calc.genoprob and/or sim.geno with
a more coarse grid. hyper <- calc.genoprob(hyper, step=10, err=0.01)
hyper <- sim.geno(hyper, step=10, n.draws=64, err=0.01) out2.hk <- scantwo(hyper, method="hk")
out2.em <- scantwo(hyper)
out2.imp <- scantwo(hyper, method="imp") The output is an object with class scantwo. The function
plot.scantwo may be used to plot the results. The
upper triangle contains LOD scores for tests of epistasis, while the
lower triangle contains LOD scores for the full model. plot(out2.hk)
plot(out2.em)
plot(out2.imp) The function summary.scantwo lists the interesting
aspects of the output. For each pair of chromosomes $(k,l)$, it
calculates the maximum LOD score for the full model, $M_f(k,l)$; a
LOD score indicating evidence for a second QTL, allowing for epistasis),
$M_fv1(k,l)$; a LOD score indicating evidence for
epistasis, $M_i(k,l)$; the LOD score for the additive QTL model,
$M_a(k,l)$; and a LOD score indicating evidence for a second QTL,
assuming no epistasis, $M_av1(k,l)$. You must provide five LOD thresholds, corresponding to the above five
LOD scores, and in that order. A chromosome pair is printed if either
(a) $M_f(k,l) >= T_f$ and ($M_fv1(k,l) >= T_fv1$ or $M_i(k,l) >= T_i$), or (b)
$M_a(k,l) >= T_a$ and $M_av1(k,l) >= T_av1$. summary(out2.em, thresholds=c(6.2, 5.0, 4.6, 4.5, 2.3))
summary(out2.em, thresholds=c(6.2, 5.0, Inf, 4.5, 2.3)) In the latter case, the interaction LOD score will be ignored. The function max.scantwo returns the maximum joint
and additive LODs for a two-dimensional genome scan. max(out2.em) Permutation tests may also performed with scantwo;
it may take a few days of CPU time. The output is a list containing the
maxima of the above five LOD scores for each of the imputations. operm2 <- scantwo(hyper, method="hk", n.perm=100)
summary(operm2, alpha=0.05)Broman, K. W., Wu, H., Sen, Ś. and Churchill, G. A. (2003) R/qtl: QTL mapping in experimental crosses. Bioinformatics 19, 889--890.
Haley, C. S. and Knott, S. A. (1992) A simple regression method for mapping quantitative trait loci in line crosses using flanking markers. Heredity 69, 315--324.
Lander, E. S. and Botstein, D. (1989) Mapping Mendelian factors underlying quantitative traits using RFLP linkage maps. Genetics 121, 185--199.
Lincoln, S. E. and Lander, E. S. (1992) Systematic detection of errors in genetic linkage data. Genomics 14, 604--610.
Sen, Ś. and Churchill, G. A. (2001) A statistical framework for quantitative trait mapping. Genetics 159, 371--387.