vignettes/yourData.Rmd
yourData.RmdIn this vignette we describe how you can use BactDating on your own data.
The first required input (let’s call it t) is a phylogenetic tree of the class phylo from the ape package, which can be loaded from a Newick or Nexus file using respectively the following commands from the ape package:
library(ape)
t=read.tree('filename.nwk')
t=read.nexus('filename.nex')This tree should have branch lengths measured in units of substitutions. In other words, the branch lengths should not be measured in substitutions per site. If you are not sure whether the branch lengths are measured correctly or not, try to compute sum(t$edge.length). This should be the total number of substitutions throughout the tree, so if you get a value below one or even only a bit above one, your branch lengths are probably not in the correct unit.
If the branch lengths are measured per site, you will need to rescale them using the command:
t$edge.length=t$edge.length*Lwhere L is the number of sites which were used to build this tree.
Alternatively, you can load directly from the output of ClonalFrameML or Gubbins using the following commands respectively:
t=loadCFML(prefix)
t=loadGubbins(prefix)This will have the advantage to account for the recombination events detected by ClonalFrameML or Gubbins when performing the dating.
The second required input (let’s call it d) is the dates at which the isolates were sampled. Dates need to be expressed decimally, for example 2015.5 means 1st July 2015. If needed, you can convert into decimal years from other date formats using the decimal_date function of the lubridate package. This input can be simply a vector of decimal dates, in which case the order needs to be the same as the order of the tree tips in t$tip.label. For example, if t$tip.label is (A,B,C) then d=c(2010,2011,2012) indicates that A was isolated in 2010, B in 2011 and C in 2012.
Alternatively, the vector d does not have to be ordered if names(d) corresponds to the tip names. For example, d=c(2011,2010,2012)with names(d)=c('B','A','C') would have the same meaning as in the previous example.
If some isolation dates are unknown, this can be indicated using NA in the vector, in which case the date is considered unknown but within the range of known dates, ie with the range return by the command range(d,na.rm=T).
Finally, the isolation dates can be specified as an interval rather than an exact value. This is useful for example if the isolation year is 2015 but with unknown month or day. To do so, d needs to be a matrix with two columns, with the first column indicating the lower bound and the second column indicating the upper bound of the interval. For example, if year is the vector of isolation years, but the exact date in each year is unknown, you can specify this by building the matrix d as follows:
d=cbind(year,year+1)Once the two inputs t and d described above are in the correct format, you are ready to run BactDating using the main command:
res=bactdate(t,d)See other vignettes for additional functionality. In particular, if your tree is unrooted you can find an optimal root using:
rooted=initRoot(t,d)Before running the main bactdate analysis, it is also a good idea to assess the strength of the temporal signal using a root-to-tip linear regression:
r=roottotip(t,d)BactDating is a MCMC algorithm which needs to be run for long enough to ensure that the results are meaningful. The length of the algorithm can be controlled using the nbIts argument of the bactdate function, which by default is 10e4 but may need increasing for example ten folds:
res=bactdate(t,d,nbIts=1e5)To test the MCMC convergence, a good start is to check that the traces of the parameters look stable using the command:
plot(res,'trace')For further testing of convergence, you can export the BactDating result to the format required by the coda package using the command:
mcmc=as.mcmc(res)You can then compute for example the effective sample size of the parameters which should be at least 100 for each paramater, using:
effectiveSize(mcmc)