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MetaCurator

Software for curating reference sequence databases used in barcoding, metabarcoding and metagenomics

MetaCurator uses HMMER (http://hmmer.org/), MAFFT (https://mafft.cbrc.jp/alignment/software/linux.html) and VSEARCH (https://github.com/torognes/vsearch) to search through sequences available from broad origions (such as NCBI Nucleotide, NCBI Genome and/or BOLD), identify and extract the precise barcode marker region of interest, discard irrelevent sequences and sequence regions, and dereplicate the resulting reference data in a taxonomically-supervised fashion.

With respect to similar softare, MetaCurator represents a flexible, generalizable and highly-sensitive alternative to the CRUX module of ANACAPA (https://github.com/limey-bean/Anacapa/) and the conserved marker curation mode of Metaxa2_dbb (https://doi.org/10.1093/bioinformatics/bty482).

Dependencies

MetaCurator is a command-line only toolkit which runs on typical Unix/Linux environments. It requires the following software to be installed and globally accessible.

Python 2.7.5 or greater
Perl 5.16.0 or compatible
VSEARCH 2.8.1 or compatible
HMMER3 3.1b2 or compatible
MAFFT 7.270 or compatible

Installation

Download and unpack tarfile of current release

wget https://github.com/RTRichar/MetaCurator/archive/v1.0.1.tar.gz
tar xzvf v1.0.1.tar.gz 

After unpacking, export the MetaCurator directory to $PATH if you're using a local machine, or add the directory path to the appropriate login configuration file if you are using a remote cluster

Test MetaCurator Installation

An example dataset of 10,000 sequences annotated as rbcL in NCBI Nucleotide is provided along with a Taxonomizr formated taxonomy file and a set of 8 hand-picked rbcL sequences manually trimmed to be representative of a commonly used barcoding region. Running the software on this test set should take approximately one hour on a typical single core machine with 4 GB of RAM.

cd MetaCurator-1.0.1/TestMetaCurator
MetaCurator.py -r rbcL_Reps.fa -i rbcL_sample.fa -it rbcL_sample.tax -tf True -ct True -of Test.fa -ot Test.tax --SaveTemp True 2> rbcL_log.txt

Usage

A general workflow for using MetaCurator is available at the following link: https://github.com/RTRichar/MetabarcodeDBsV2/blob/master/Workflow.md

  • if you're running MetaCurator on a fasta file with sequences that are split over multiple lines (such as the default NCBI download format), you'll need to concatenate lintes into continuous sequences as decribed through the link above

Citation and further information

Richardson, RT, DB Sponsler, H McMinn-Sauder & RM Johnson. (2019). MetaCurator: A hidden Markov model-based toolkit for extracting and curating sequences from taxonomically-informative genetic markers. Methods in Ecology and Evolution. https://doi.org/10.1111/2041-210X.13314