WASTER is a coalesence-aware de novo species tree inference tool, which means it can take as inputs raw reads in FASTQ format. Paticularly, WASTER can accurately infer species tree even from Illumina reads with only 1.5X depth. WASTER infers the species tree by first calling SNPs from reads/assembies and then invoking CASTER to reconstruct the species tree from the SNPs.
TBA
Many ASTER tools have been integrated in PhyloSuite, an integrated and scalable desktop platform for streamlined molecular sequence data management and evolutionary phylogenetics studies.
Please check out our software with GUI. Simply download the zip file, extract the contents, enter exe
folder, and click aster-gui.exe
.
Contact [email protected]
, [email protected]
, or post on ASTER issues page.
- The rest of this TUTORIAL file
- Forums (feel free to ask questions or ask for help running ASTER):
- User group discussions
- ASTER issues page
- QQ group: 130635706
For most users, installing ASTER is very easy! Download using one of two approaches:
- You simply need to download the zip file for Windows/MacOS/Linux and extract the contents to a folder of your choice.
- Alternatively, you can clone the github repository and checkout the branch named Windows/MacOS/Linux.
Binary files should be in the exe
folder for Windows or bin
folder otherwise. If you are lucky, these may just work as is and you may not need to build at all.
- In terminal,
cd
into the downloaded directory and runmake
.
- If you see
*** Installation complete! ***
then you are done! - If you see
Command 'g++' not found
then before rerunningmake
,- Debian (Ubuntu) users try
sudo apt update sudo apt install g++
- CentOS (RedHat) users try
sudo yum update sudo yum install gcc-c++
- Unix (MacOS) users should be prompted for installing
g++
and please click "install". If no prompt, tryg++
.
- Debian (Ubuntu) users try
- If you see "error" when running
make
, please trymake waster-site
instead and file a bug report.
- Binary files should be in the
bin
folder.
- Executables for x86-64 are available in
exe
folder and it is very likely that they already work. - Windows Subsystem for Linux (WSL) is HIGHLY recommanded if you need to install on your own! Please follow instructions in "For Linux/Unix/WSL users" section.
- To compile windows excutables:
- Download MinGW and install posix version for your architecture (eg. x86-64)
- Add path to
bin
folder of MinGW to system environment variablePATH
- Double click
make.bat
inside the downloaded directory
Please check out our software with GUI. Simply download the zip file, extract the contents, enter exe
folder, and click aster-gui.exe
.
If you have overlapping paired-end sequencing reads, please make sure you have merged them using BBMerge
or alike.
This will improve the accuracy of WASTER on biological datasets!
If you have non-overlapping paired-end sequencing reads, great!
With the same sequencing depth, WASTER prefers non-overlapping reads.
The input is a text file containing a list of species names each followed by a Fasta/Fastq file (one species name and one file per line). For example:
speciesA example/waster/filename1.fa
speciesB example/waster/filename2.fa
speciesC example/waster/filename3.fq
speciesD example/waster/filename4.fq
Make sure input files are not zipped.
When multiple individuals from the same species are available, you need to let WASTER to know that they are from the same species.
- You can give multiple individuals from the same species the same name in the input list.
speciesA example/waster/filename1.fa
speciesA example/waster/filename2.fa
speciesB example/waster/filename3.fq
speciesB example/waster/filename4.fq
...
WARNING: Never use this method to input pair-end reads files!!!
If your pair-end reads overlap, then merge them using BBMerge
or alike; otherwise cat
them into one file.
The output in is Newick format and gives:
- the species tree topology
- branch supports measured as local bootstrap support (>95.0 means good)
- It can also annotate branches with other quantities, such as quartet scores and local bootstraps for all three topologies.
ASTER currently has no GUI. You need to run it through the command-line. In a terminal/PowerShell, go to the directory (location) where you have downloaded ASTER and issue the following command:
bin/waster-site
This will give you a list of options available. If you are using Windows, please replace bin/waster-site
with .\exe\waster-site.exe
.
To find the species tree with input from in a file called INPUT_FILE
, use:
bin/waster-site INPUT_FILE
or
bin/waster-site -i INPUT_FILE
In the first case, INPUT_FILE is hard-coded to be the last argument for backward compatibility.
For example if you want to run waster-site
with input example/waster/input_list.txt
, then run
bin/waster-site example/waster/input_list.txt
or
bin/waster-site -i example/waster/input_list.txt
The results will be outputted to the standard output. To save the results in a file use the -o OUTPUT_FILE
option before INPUT_FILE
(Strongly recommended):
bin/waster-site -o OUTPUT_FILE INPUT_FILE
or
bin/waster-site -i INPUT_FILE -o OUTPUT_FILE
With -i INPUT_FILE
option, the order does not matter anymore. For brevity, from here on we will not demonstrate -i INPUT_FILE
cases.
To save the logs (also recommended), run:
bin/waster-site -o OUTPUT_FILE INPUT_FILE 2>LOG_FILE
For example, you can run
bin/waster-site -o example/waster/input_list.txt.stree example/waster/input_list.txt 2>example/waster/input_list.txt.log
ASTER supports multi-threading. To run program with 4 threads, add -t 4
before INPUT_FILE
:
bin/waster-site -t 4 -o OUTPUT_FILE INPUT_FILE 2>LOG_FILE
ASTER has very good parrallel efficiency up to 64 cores when input data is large. In fact, it often experiences super-linear speedup with 16 cores or more. So feel free to use as many cores as you want.
ASTER also allows rooting at an given outgroup:
bin/waster-site --root YOUR_OUTGROUP INPUT_FILE
Notice: By default, WASTER will create a 32 GB look-up table to find SNPs. So, if you are running on a machine with <64 GB memory, you need to shrink the look-up table size using -k 8
or -k 7
even for the example run.
Using -k 8
requires about 4 GB memory and -k 7
only requires <1 GB memory.
Try the following example run:
bin/waster-site -k 7 example/waster/input_list.txt
ASTER algorithm first performs R
(4 by default) rounds of search and then repeatedly performs S
(4 by default) rounds of subsampling and exploration until no improvement found.
bin/waster-site -r R -s S -o OUTPUT_FILE INPUT_FILE 2>LOG_FILE
If you want to run with more rounds of placement for ensured optimality, then you can run with
bin/waster-site -r 16 -s 16 -o OUTPUT_FILE INPUT_FILE 2>LOG_FILE
or simply
bin/waster-site -R -o OUTPUT_FILE INPUT_FILE 2>LOG_FILE
If you want to place taxa on an existing fully resolved species tree, you can use -c SPECIES_TREE_IN_NEWICK_FORMAT
before INPUT_FILE
:
bin/waster-site -o OUTPUT_FILE -c SPECIES_TREE_IN_NEWICK_FORMAT INPUT_FILE
Specifically, you can score and annotate a fully resolved species tree containing all taxa with -c SPECIES_TREE_IN_NEWICK_FORMAT
. If want to score a species tree or you want to place only one taxon onto the tree, you can use
bin/waster-site -r 1 -s 0 -o OUTPUT_FILE -c SPECIES_TREE_IN_NEWICK_FORMAT INPUT_FILE
or simply,
bin/waster-site -C -o OUTPUT_FILE -c SPECIES_TREE_IN_NEWICK_FORMAT INPUT_FILE
If you want to give hints by providing candidate species trees or trees similar to the species tree, you can use -g SPECIES_TREES_IN_NEWICK_FORMAT
before INPUT_FILE
:
bin/waster-site -o OUTPUT_FILE -g SPECIES_TREES_IN_NEWICK_FORMAT INPUT_FILE