knitr::opts_chunk$set( collapse = TRUE, comment = "#>", fig.width=4.5, fig.height=3.5 )
We demonstrate how to simulate NGS data under various genotype distributions, then fit these data using flexdog
. The genotyping methods are described in Gerard et al. (2018).
Let's suppose that we have 100 hexaploid individuals, with varying levels of read-depth.
set.seed(1) library(updog) nind <- 100 ploidy <- 6 sizevec <- round(stats::runif(n = nind, min = 50, max = 200))
We can simulate their read-counts under various genotype distributions, allele biases, overdispersions, and sequencing error rates using the rgeno
and rflexdog
functions.
Suppose these individuals are all siblings where the first parent has 4 copies of the reference allele and the second parent has 5 copies of the reference allele. Then the following code, using rgeno
, will simulate the individuals' genotypes.
true_geno <- rgeno(n = nind, ploidy = ploidy, model = "f1", p1geno = 4, p2geno = 5)
Once we have their genotypes, we can simulate their read-counts using rflexdog
. Let's suppose that there is a moderate level of allelic bias (0.7) and a small level of overdispersion (0.005). Generally, in the real data that I've seen, the bias will range between 0.5 and 2 and the overdispersion will range between 0 and 0.02, with only a few extremely overdispersed SNPs above 0.02.
refvec <- rflexdog(sizevec = sizevec, geno = true_geno, ploidy = ploidy, seq = 0.001, bias = 0.7, od = 0.005)
When we plot the data, it looks realistic
plot_geno(refvec = refvec, sizevec = sizevec, ploidy = ploidy, bias = 0.7, seq = 0.001, geno = true_geno)
We can test flexdog
on these data
fout <- flexdog(refvec = refvec, sizevec = sizevec, ploidy = ploidy, model = "f1")
flexdog
gives us reasonable genotyping, and it accurately estimates the proportion of individuals mis-genotyped.
plot(fout) ## Estimated proportion misgenotyped fout$prop_mis ## Actual proportion misgenotyped mean(fout$geno != true_geno)
Now run the same simulations assuming the individuals are in Hardy-Weinberg population with an allele frequency of 0.75.
true_geno <- rgeno(n = nind, ploidy = ploidy, model = "hw", allele_freq = 0.75) refvec <- rflexdog(sizevec = sizevec, geno = true_geno, ploidy = ploidy, seq = 0.001, bias = 0.7, od = 0.005) fout <- flexdog(refvec = refvec, sizevec = sizevec, ploidy = ploidy, model = "hw") plot(fout) ## Estimated proportion misgenotyped fout$prop_mis ## Actual proportion misgenotyped mean(fout$geno != true_geno) ## Estimated allele frequency close to true allele frequency fout$par$alpha
Gerard, David, Luís Felipe Ventorim Ferrão, Antonio Augusto Franco Garcia, and Matthew Stephens. 2018. "Genotyping Polyploids from Messy Sequencing Data." Genetics 210 (3). Genetics: 789–807. https://doi.org/10.1534/genetics.118.301468.
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