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Wolbachia MLST protocols

Following is a quick guideline of the steps necessary for typing a Wolbachia strain. Please refer to the protocols below for detailed information.

Step flowchart

  1. Choose the sample of strains you wish to type.
  2. Download the Isolates Form (below), gather all possible information about each strain and start filling the form.
  3. For each strain, sequence the five MLST genes using primers and protocol provided below (protocol #1 or #2 for single or double AB infected species, respectively). Sequencing of wsp is not required, but strongly suggested (see the wsp database for primers and protocol).
  4. Prepare sequences in FASTA format using the allele templates (below) as reference (for details on this and the following steps please refer to protocol #3 below).
  5. Query the Profiles database for allele assignment of all the your MLST sequences.
  6. Send all new alleles to the curator using the automated submission system.
  7. Complete the Isolates Form with the allelic profiles.
  8. Query each allelic profile to the Profiles database.
  9. Download the Allelic profiles form (below) and fill it with the new allelic profiles.
  10. Send the following files to the curator using the automated submission system.
    • Complete Allelic profiles form (containing only the new allelic profiles discovered)
    • Complete Isolates form

The protocols below contain detailed information on the experimental conditions and procedures of the steps listed above.

  1. Amplification and sequencing of single infected individuals
  2. Amplification and sequencing of double AB infected individuals
  3. Preparation and submission of data
  4. Allele templates

Citing the database

The preferred format for citing this website in publications is:

This publication made use of the MLST website (https://pubmlst.org/ wolbachia/) sited at the University of Oxford (Jolley & Maiden 2010, BMC Bioinformatics, 11:593). The development and maintenance of this site has been funded by the Wellcome Trust.

Citation for use of the Wolbachia MLST system and database is Baldo et al. 2006, Appl Environ Microbiol 72:7098-7110. Development and curation of the Wolbachia MLST system has been supported by funds from the US National Science Foundation to Jack Werren as part of community outreach activities.


Sequence database
Sequences: 3,673
Profiles (MLST): 576
Last updated: 2020-02-24

Isolate database
Isolates: 2,019
Last updated: 2020-02-24