Ipyrad

Introduction

The software ipyrad is an interactive toolkit for assembly and analysis of restriction-site associated genomic data sets (e.g., RAD, ddRAD, GBS) for population genetic and phylogenetic studies. [1]

At the moment, there is no central installation of ipyrad, however, you can easily install it yourself using Anaconda3 as described below.

Installation

To install ipyrad you first need to load a module for Anaconda3. In this example, we use Anaconda3/2020.02 which can be found in hpc-env/8.3 (if you want to use a different version/environment you can search with module av Anaconda3 or module spider Anaconda3):

[carl]$ module load hpc-env/8.3
[carl]$ module load Anaconda/2020.02

The next step is to create a new environment for ipyrad with the command:

[carl]$ conda create --name ipyrad
Collecting package metadata (current_repodata.json): done
Solving environment: done

## Package Plan ##
  environment location: /user/abcd1234/.conda/envs/ipyrad

Proceed ([y]/n)?

Preparing transaction: done
Verifying transaction: done
Executing transaction: done

The name for the environment can be freely chosen and it will be created after you have confirmed to proceed with pressing (y and)enter. You may see a warning about an outdated conda which you can safely ignore (or, if you wish, you can switch to a newer module of Anaconda3 if available).

The new environment can now be activated. We recommend using the command(*):

[carl]$ source activate ipyrad
(ipyrad) [carl]$

You will notice the change of the command-line prompt to indicate the active environment. Packages that are now installed with conda install will be installed in this environment and not interfere with other software installations.

(*) The alternative conda activate requires you to use the command conda init bash first which modifies your .bashrc and more or less forces you to always use the same version of Anaconda3.

Now you can install ipyrad along with a the package mpi4py for parallel computing?

(ipyrad) [carl]$ conda install ipyrad -c bioconda
(ipyrad) [carl]$ conda install mpi4py -c conda-forge

These commands will take a moment to complete but after that ipyrad is ready to use. And next time you log in or in a job script you only need the commands

[carl]$ module load hpc-env/8.3
[carl]$ module load Anaconda/2020.02
[carl]$ source activate ipyrad

to get started. If you want to leave the environment you can always type

(ipyrad) [carl]$ conda deactivate

which should return you to the normal command-line prompt.

Using ipyrad on CARL

Preparations and First Tests

Following the Introductory tutorial to the CLI you can start by downloading some test data and creating a parameter file in a new directory under $WORK:

(ipyrad) [carl]$ mkdir $WORK/ipyrad_test
(ipyrad) [carl]$ cd $WORK/ipyrad_test
(ipyrad) [carl]$ curl -LkO https://eaton-lab.org/data/ipsimdata.tar.gz
  % Total    % Received % Xferd  Average Speed   Time    Time     Time  Current
                                 Dload  Upload   Total   Spent    Left  Speed 
100 11.8M  100 11.8M    0     0  8514k      0  0:00:01  0:00:01 --:--:-- 8508k 
(ipyrad) [carl]$ tar -xzf ipsimdata.tar.gz
(ipyrad) [carl]$ ipyrad -n iptest
  New file 'params-iptest.txt' created in /gss/work/lees4820/ipyrad

The resulting file params-iptest.txt has to be opened in a text editor to add the locations of the raw non-demultiplexed fastq file and the barcodes file. With the test data the first couple of lines should look like this:

(ipyrad) [carl]$ $ head params-iptest.txt
------- ipyrad params file (v.0.9.53)-------------------------------------------
iptest                         ## [0] [assembly_name]: Assembly name. Used to name output directories for assembly steps
/gss/work/abcd1234/ipyrad      ## [1] [project_dir]: Project dir (made in curdir if not present)
./ipsimdata/rad_example_R1_.fastq.gz      ## [2] [raw_fastq_path]: Location of raw non-demultiplexed fastq files
./ipsimdata/rad_example_barcodes.txt      ## [3] [barcodes_path]: Location of barcodes file
                               ## [4] [sorted_fastq_path]: Location of demultiplexed/sorted fastq files
denovo                         ## [5] [assembly_method]: Assembly method (denovo, reference)

A simple test can be performed with this command:

(ipyrad) [carl]$ ipyrad -p params-iptest.txt -s 1 -c 8
 -------------------------------------------------------------
  ipyrad [v.0.9.53]
  Interactive assembly and analysis of RAD-seq data
 -------------------------------------------------------------
  Parallel connection | hpcl004: 8 cores

  Step 1: Demultiplexing fastq data to Samples

The program performs the first step of a work flow (-s 1) using a total of 8 cores (-c 8) on the login node. We can now remove the newly created data files and directories with

(ipyrad) [carl]$ rm -r iptest_fastqs/ iptest.json

to avoid error message in the next steps.