Monday, November 23, 2015

Find the best model of evolution for Trpc2 using modelomatic:
#For this analysis, all stop codons need to be removed.
#For troubleshooting modelOmatic, I wrote a more detailed post here.
#Navigate to the correct directory and run this command:
modelomatic_OSX Trpc2_alignment_noStops.phy bionj Trpc2_modelomatic_normal.txt 0 normal

Interestingly REV+4dG is the best model, despite it being a protein coding gene. This is essentially "GTR+G". The 4dG is showing the four rate categories (this is an arbitrary number) for the gamma parameter.

Find the "best" (ML) tree in GALI using search replicates:
#run this command
garli Trpc2_garli.conf
#Note how to implement GTR+G in GARLI. See here for more examples of other models.
#Here is my configuration file (Trpc2_garli.conf):
[general]
datafname = Trpc2_garli.nex
constraintfile = none
streefname = stepwise
attachmentspertaxon = 50
ofprefix = Trpc2.GTR4DG
randseed = -1
availablememory = 512
logevery = 10
saveevery = 100
refinestart = 1
outputeachbettertopology = 0
outputcurrentbesttopology = 0
enforcetermconditions = 1
genthreshfortopoterm = 20000
scorethreshforterm = 0.05
significanttopochange = 0.01
outputphyliptree = 0
outputmostlyuselessfiles = 0
writecheckpoints = 0
restart = 0
outgroup = 1
resampleproportion = 1.0
inferinternalstateprobs = 0
outputsitelikelihoods = 0
optimizeinputonly = 0
collapsebranches = 1

searchreps = 8
bootstrapreps = 0

[model1]
datatype = nucleotide
ratematrix = 6rate
statefrequencies = estimate
ratehetmodel = gamma
numratecats = 4
invariantsites = none

[master]
nindivs = 4
holdover = 1
selectionintensity = 0.5
holdoverpenalty = 0
stopgen = 5000000
stoptime = 5000000

startoptprec = 0.5
minoptprec = 0.01
numberofprecreductions = 10
treerejectionthreshold = 50.0
topoweight = 1.0
modweight = 0.05
brlenweight = 0.2
randnniweight = 0.1
randsprweight = 0.3
limsprweight =  0.6
intervallength = 100
intervalstostore = 5
limsprrange = 6
meanbrlenmuts = 5
gammashapebrlen = 1000
gammashapemodel = 1000
uniqueswapbias = 0.1
distanceswapbias = 1.0

Putting bootstrap values on GARLI:
You can bootstrap in GARLI, but to "put" the bootstraps on the nodes, you have to summarize the bootstrap results using an outside method. SumTrees.py in the Dendropy python package works well. I basically followed the exact instructions somebody wrote here.

Running bootstrap iterations in GARLI:
#I ran four independent runs simultaneously of 250 bootstrap iterations that will combine to = 1000.
#Here is my GARLI configuration file:
[general]
datafname = Trpc2_garli.nex
constraintfile = none
streefname = stepwise
attachmentspertaxon = 50
ofprefix = Trpc2.GTR4DG.boot
randseed = -1
availablememory = 512
logevery = 10
saveevery = 100
refinestart = 1
outputeachbettertopology = 0
outputcurrentbesttopology = 0
enforcetermconditions = 1
genthreshfortopoterm = 20000
scorethreshforterm = 0.05
significanttopochange = 0.01
outputphyliptree = 0
outputmostlyuselessfiles = 0
writecheckpoints = 0
restart = 0
outgroup = 1
resampleproportion = 1.0
inferinternalstateprobs = 0
outputsitelikelihoods = 0
optimizeinputonly = 0
collapsebranches = 1

searchreps = 1
bootstrapreps = 250

[model1]
datatype = nucleotide
ratematrix = 6rate
statefrequencies = estimate
ratehetmodel = gamma
numratecats = 4
invariantsites = none

[master]
nindivs = 4
holdover = 1
selectionintensity = 0.5
holdoverpenalty = 0
stopgen = 5000000
stoptime = 5000000

startoptprec = 0.5
minoptprec = 0.01
numberofprecreductions = 10
treerejectionthreshold = 20.0
topoweight = 1.0
modweight = 0.05
brlenweight = 0.2
randnniweight = 0.1
randsprweight = 0.3
limsprweight =  0.6
intervallength = 100
intervalstostore = 5
limsprrange = 6
meanbrlenmuts = 5
gammashapebrlen = 1000
gammashapemodel = 1000
uniqueswapbias = 0.1
distanceswapbias = 1.0

Installing dendropy
#We have "pip" set up on our server, making the python package easy to install:
sudo pip install -U dendropy

Running SumTrees
#place all of your bootstrap outputs and your best tree from GARLI in its own folder

cd /Volumes/Yango/Trpc2/garli/

mkdir sumtrees
cp Trpc2.GTR4DG.best.tre sumtrees/
cp Trpc2.GTR4DG.boot.boot.tre sumtrees/
cp Trpc2.GTR4DG.boot2.boot.tre sumtrees/
cp Trpc2.GTR4DG.boot3.boot.tre sumtrees/
cp Trpc2.GTR4DG.boot4.boot.tre sumtrees/

#from within the folder, run sumtrees.py
#your target tree is the "best" tree from your GARLI output

sumtrees.py --decimals=0 --percentages --no-analysis-metainformation --no-taxa-block Trpc2.GTR4DG.boot.boot.tre Trpc2.GTR4DG.boot2.boot.tre Trpc2.GTR4DG.boot3.boot.tre Trpc2.GTR4DG.boot4.boot.tre --target=Trpc2.GTR4DG.best.tre --output=supportOnBest.phy

Viewing tree with bootstraps
Now, open supportOnBest.phy in FigTree. Click on "node labels" and select "support" as the node label. These are your bootstrap values.

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