GenomeTrakr WGS Protocol Collection and Workflow for MiSeq

Ruth Timme, Maria Balkey, Julie Haendiges, Candace Hope Bias, Tina Lusk Pfefer

Published: 2024-03-22 DOI: 10.17504/protocols.io.3byl4bwyjvo5/v3

Disclaimer

Abstract

Here we have created a collection of all the protocols used for WGS using the MiSeq, in order, from sample extraction to NCBI submission.

This collection has three sections:

  • Section 1: WGS Wet lab workflow for Illumina MiSeq
  • Section 2: Dry lab workflow for sequence QC and NCBI submission - Direct Submission
  • Section 3: Dry lab workflow for sequence QC and NCBI submission - PulseNet labs

Version history:

V3:

  • Links to protocols updated with latest version,
  • Added new protocol: DNA Extraction from Inoculated Broth for WGS using the Maxwell RSC Cultured Cells DNA Kit,
  • Added new protocol: Querying for Bacterial Pathogen Genomic Data at NCBI.

Steps

WGS Wet Lab Workflow for Illumina MiSeq

Dry Lab Workflow for Sequence QC and NCBI Submission - Direct Submission

6.

The following protocols are also included in a Springer Methods book chapter collection: Utilizing the Public GenomeTrakr Database for Foodborne Pathogen Traceback (protocols.io)

9.

Submit sequence and metadata to NCBI:

NCBI submission protocol for microbial pathogen surveillance

11.

Dry Lab Workflow for Sequence QC and NCBI Submission - PulseNet Labs

12.

Check sequence quality:

Follow the CDC's guidance for assessing QC in BioNumerics

13.

Populate NCBI template:

***NOTE: For NCBI submission for fields not included in the BioNumerics Template, please remember to include the name of the laboratory sequencing the isolates and the surveillance effort name in the sequenced_by and project_name fields as described in Step 5 of this protocol.

Populating NCBI template for submissions using BioNumerics

14.

Submit sequence and metadata to NCBI:

Follow the CDC's guidance for NCBI submissions through BioNumerics.

15.

Query data at NCBI

16.

Update, retract, or replace these records in NCBI databases, if necessary:

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