From 832f6c20533062af20802a3d180526b010ef7dd6 Mon Sep 17 00:00:00 2001
From: Alexey Kozlov
diff --git a/doc/CellPhy-Tutorial.Rmd b/doc/CellPhy-Tutorial.Rmd
index 5cece98..af0ffae 100644
--- a/doc/CellPhy-Tutorial.Rmd
+++ b/doc/CellPhy-Tutorial.Rmd
@@ -1,7 +1,7 @@
---
title: "CellPhy - The hard way (tree search, bootstrapping, mutation mapping)"
author: 'Alexey Kozlov, [João M Alves*](mailto:jmfernandesalves@gmail.com), Alexandros Stamatakis & David Posada'
-date: "July 2020"
+date: "March 2021"
output:
pdf_document:
number_sections: yes
@@ -59,7 +59,7 @@ We will run all commands from the `example` directory where sample input files r
* 1) matrix of genotypes in FASTA or **PHYLIP** format - _CRC24.ToySet.phy_
* 2) [**standard VCF file**](https://samtools.github.io/hts-specs/VCFv4.3.pdf) - _CRC24.ToySet.vcf_
-When the input is a genotype matrix, genotypes are encoded as shown in Table S2 of Kozlov _et al._ 2020. When the input is a VCF, CellPhy can be run in two distinct modes. The first mode (“**CellPhy-EP17**”) requires a VCF with at least the GT field (that stores the genotype calls), in which case CellPhy simply extracts the genotype matrix. The second mode (“**CellPhy-GL**”) requires a VCF with the PL field (which stores the phred-scaled genotype likelihoods) and uses instead the likelihoods of each genotype.
+When the input is a genotype matrix, genotypes are encoded as shown in Table S2 of Kozlov _et al._ 2020. When the input is a VCF, CellPhy can be run in two distinct modes. The first mode (“**CellPhy-ML**”) requires a VCF with at least the GT field (that stores the genotype calls), in which case CellPhy simply extracts the genotype matrix. The second mode (“**CellPhy-GL**”) requires a VCF with the PL field (which stores the phred-scaled genotype likelihoods) and uses instead the likelihoods of each genotype.
Here's how they look like:
Let us now infer a tree with default parameters using both models of CellPhy. Here, we will use a single thread and set a fixed random seed to ensure reproducibility.
-## **Tree inference with "CellPhy-EP17"** -This model will take as input a genotype matrix to infer a tree under the genotype error model "_GTGTR4+FO+E_". If the input file is a PHYLIP file, CellPhy will automatically recognize the file type and proceed with the analysis. In contrast, if the input is a VCF file we need to set `--prob-msa off` which will instruct CellPhy to use the genotype field (GT) instead of the genotype likelihoods (PL). +## **Tree inference with "CellPhy-ML"** +
This model will take as input a genotype matrix to infer a tree under the genotype error model "_GT16+FO+E_". If the input file is a PHYLIP file, CellPhy will automatically recognize the file type and proceed with the analysis. In contrast, if the input is a VCF file we need to set `--prob-msa off` which will instruct CellPhy to use the genotype field (GT) instead of the genotype likelihoods (PL). These two runs should therefore return exactly the same result:
\scriptsize ```{r, engine = 'bash', eval = FALSE} -$ ../cellphy.sh RAXML --msa CRC24.ToySet.phy --model GTGTR4+FO+E --seed 2 --threads 1 --prefix CRC24.PHY.EP17.Tree -$ ../cellphy.sh RAXML --msa CRC24.ToySet.vcf --model GTGTR4+FO+E --seed 2 --threads 1 --prefix CRC24.VCF.EP17.Tree --prob-msa off +$ ../cellphy.sh RAXML --msa CRC24.ToySet.phy --model GT16+FO+E --seed 2 --threads 1 --prefix CRC24.PHY.ML16.Tree +$ ../cellphy.sh RAXML --msa CRC24.ToySet.vcf --model GT16+FO+E --seed 2 --threads 1 --prefix CRC24.VCF.ML16.Tree --prob-msa off ``` \normalsize @@ -126,12 +126,12 @@ Analysis options: Optimized model parameters: Partition 0: noname - Error model (ML): P17, SEQ_ERROR: 0.000000, ADO_RATE: 0.698852 + Error model (ML): P20, SEQ_ERROR: 0.000000, ADO_RATE: 0.698807 Rate heterogeneity: NONE - Base frequencies (ML): 0.060768 0.054332 0.087159 0.066360 0.093350 0.244303 0.026375 0.019954 0.255222 0.092177 - Substitution rates (ML): 17.783518 17.783518 17.783518 66.024749 1000.000000 0.001000 17.783518 17.783518 17.783518 [...] + Base frequencies (ML): 0.046975 0.076605 0.093862 0.059652 0.000047 0.000047 0.062979 0.000047 0.000047 0.000047 [...] + Substitution rates (ML): 0.001000 0.001000 0.001000 27.354070 989.178276 93.810669 0.001000 0.001000 0.001000 27.354070 [...] -Final LogLikelihood: -9731.527620 +Final LogLikelihood: -9536.825816 ``` @@ -142,9 +142,9 @@ Final LogLikelihood: -9731.527620 \scriptsize ```{r, engine = 'bash', eval = FALSE} -$ grep "Final LogLikelihood:" CRC24.{PHY,VCF}.EP17.Tree.raxml.log -CRC24.PHY.EP17.Tree.raxml.log:Final LogLikelihood: -9731.527620 -CRC24.VCF.EP17.Tree.raxml.log:Final LogLikelihood: -9731.527620 +$ grep "Final LogLikelihood:" CRC24.{PHY,VCF}.ML16.Tree.raxml.log +CRC24.PHY.ML16.Tree.raxml.log:Final LogLikelihood: -9536.825816 +CRC24.VCF.ML16.Tree.raxml.log:Final LogLikelihood: -9536.825816 ``` \normalsize @@ -155,7 +155,7 @@ CRC24.VCF.EP17.Tree.raxml.log:Final LogLikelihood: -9731.527620 \scriptsize ```{r, engine = 'bash', eval = FALSE} -$ ../cellphy.sh RAXML --msa CRC24.ToySet.vcf --model GTGTR4+FO --seed 2 --threads 1 --prefix CRC24.VCF.GL.Tree +$ ../cellphy.sh RAXML --msa CRC24.ToySet.vcf --model GT16+FO --seed 2 --threads 1 --prefix CRC24.VCF.GL16.Tree ``` \normalsize @@ -170,20 +170,45 @@ Optimized model parameters: Partition 0: noname Rate heterogeneity: NONE - Base frequencies (ML): 0.139898 0.190237 0.201048 0.162514 0.047442 0.102149 0.011684 0.007876 0.093886 0.043266 - Substitution rates (ML): 0.140301 0.140301 0.140301 523.825038 686.075522 74.470843 0.140301 0.140301 0.140301 [...] + Base frequencies (ML): 0.123626 0.175592 0.186970 0.142850 0.000124 0.000124 0.027265 0.013460 0.000124 0.000124 [...] + Substitution rates (ML): 0.001000 0.001000 0.001000 304.877289 263.121312 306.737332 0.001000 0.001000 0.001000 [...] -Final LogLikelihood: -30265.905808 +Final LogLikelihood: -30087.656427 [...] -Best ML tree saved to: CRC24.VCF.GL.Tree.raxml.bestTree -All ML trees saved to: CRC24.VCF.GL.Tree.raxml.mlTrees -Optimized model saved to: CRC24.VCF.GL.Tree.raxml.bestModel +Best ML tree saved to: CRC24.VCF.GL16.Tree.raxml.bestTree +All ML trees saved to: CRC24.VCF.GL16.Tree.raxml.mlTrees +Optimized model saved to: CRC24.VCF.GL16.Tree.raxml.bestModel ``` \normalsize +## **Tree inference with the GT10 model** + +In addition to the default genotype model with 16 states (_GT16_), CellPhy offers a faster approximate model with 10 states -- _GT10_: + +\scriptsize + +```{r, engine = 'bash', eval = FALSE} +$ ../cellphy.sh RAXML --msa CRC24.ToySet.phy --model GT10+FO+E --seed 2 --threads 1 --prefix CRC24.PHY.ML10.Tree +$ ../cellphy.sh RAXML --msa CRC24.ToySet.vcf --model GT10+FO --seed 2 --threads 1 --prefix CRC24.VCF.GL10.Tree + +``` +\normalsize + +On unphased genotype input data, which in fact only contain 10 states, the _GT10_ model appears to be as accurate as _GT16_, but requires only half of the time: + +\scriptsize + +```{r, engine = 'bash', eval = FALSE} +$ grep Elapsed CRC24.VCF.ML1?.Tree.raxml.log +CRC24.VCF.ML10.Tree.raxml.log:Elapsed time: 122.266 seconds +CRC24.VCF.ML16.Tree.raxml.log:Elapsed time: 240.350 seconds +``` + +\normalsize + *** # **Bootstrapping trees and tree branch support** @@ -193,8 +218,8 @@ Optimized model saved to: CRC24.VCF.GL.Tree.raxml.bestModel \scriptsize ```{r, engine = 'bash', eval = FALSE} -$ ../cellphy.sh RAXML --bootstrap --msa CRC24.ToySet.vcf --model GTGTR4+FO --seed 2 --threads 1 --bs-trees 100 \ - --prefix CRC24.VCF.GL.Boot +$ ../cellphy.sh RAXML --bootstrap --msa CRC24.ToySet.vcf --model GT16+FO --seed 2 --threads 1 --bs-trees 100 \ + --prefix CRC24.VCF.GL16.Boot [...] [00:00:00] Data distribution: max. partitions/sites/weight per thread: 1 / 500 / 5000 @@ -205,7 +230,7 @@ $ ../cellphy.sh RAXML --bootstrap --msa CRC24.ToySet.vcf --model GTGTR4+FO --see [00:15:07] Bootstrap tree #99, logLikelihood: -29931.390293 [00:15:18] Bootstrap tree #100, logLikelihood: -29263.239019 -Bootstrap trees saved to: CRC24.VCF.GL.Boot.raxml.bootstraps +Bootstrap trees saved to: CRC24.VCF.GL16.Boot.raxml.bootstraps ``` \normalsize @@ -214,9 +239,9 @@ Bootstrap trees saved to: CRC24.VCF.GL.Boot.raxml.bootstraps \scriptsize ```{r, engine = 'bash', eval = FALSE} -$ ../cellphy.sh RAXML --support -tree CRC24.VCF.GL.Tree.raxml.bestTree \ - --bs-trees CRC24.VCF.GL.Boot.raxml.bootstraps \ - --prefix CRC24.VCF.GL.Support --threads 1 +$ ../cellphy.sh RAXML --support -tree CRC24.VCF.GL16.Tree.raxml.bestTree \ + --bs-trees CRC24.VCF.GL16.Boot.raxml.bootstraps \ + --prefix CRC24.VCF.GL16.Support --threads 1 Reading reference tree from file: CRC24.VCF.GL.Tree.raxml.bestTree Reference tree size: 25 @@ -224,7 +249,7 @@ Reference tree size: 25 Reading bootstrap trees from file: CRC24.VCF.GL.Boot.raxml.bootstraps Loaded 100 trees with 25 taxa. -Best ML tree with Felsenstein bootstrap (FBP) support values saved to: CRC24.VCF.GL.Support.raxml.support +Best ML tree with Felsenstein bootstrap (FBP) support values saved to: CRC24.VCF.GL16.Support.raxml.support ``` \normalsize @@ -235,7 +260,7 @@ Best ML tree with Felsenstein bootstrap (FBP) support values saved to: CRC24.VCF ```{r, engine = 'bash', eval = FALSE} -$ ../script/support-map.R CRC24.VCF.GL.Support.raxml.support Healthy +$ ../script/support-map.R CRC24.VCF.GL16.Support.raxml.support Healthy Generating support tree plot... Done! @@ -243,7 +268,7 @@ Done! \normalsize -This call will generate a tree plot in PDF (_CRC24.VCF.GL.Support.raxml.support.pdf_) and SVG (_CRC24.VCF.GL.Support.raxml.support.svg_) formats. The result is shown in Figure 1. +This call will generate a tree plot in PDF (_CRC24.VCF.GL16.Support.raxml.support.pdf_) and SVG (_CRC24.VCF.GL16.Support.raxml.support.svg_) formats. 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