英国上市公司365

    GenomeTrakr WGS Protocol Collection and Workflow for MiSeq

    Ruth Timme, Julie Haendiges, Maria Balkey, Tina Lusk Pfefer

    Published: 2022-05-12 DOI: 10.17504/protocols.io.3byl4bwyjvo5/v1
    AI 解读
    浏览 1528

    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

    Associated protocols:

    • Querying the NCBI database for GenomeTrakr data (protocols.io)
    • NCBI data curation protocol - SOP for editing GenomeTrakr submissions (protocols.io)

    Steps

    Wet lab

    1.

    WGS Wet lab workflow for Illumina MiSeq

    2.

    DNA Extraction

    Manual DNA Extraction using Qiagen DNeasy Blood and Tissue Kit

    3.

    DNA Quantification

    DNA Quantification using the Qubit Fluorometer

    4.

    Library Preparation

    Illumina DNA Prep (M) Tagmentation Library Preparation for use on an Illumina MiSeq Sequencer

    5.

    Sequencing

    Procedure for Operation and Maintenance of the Illumina MiSeq for Whole Genome Sequencing

    Dry lab - Direct Submission

    6.

    Dry lab workflow for sequence QC and NCBI submission - Direct Submission:

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

    7.

    Check sequence quality:

    Quality control assessment for microbial genomes: GalaxyTrakr MicroRunQC workflow

    8.

    Populate BioSample AND SRA metadata templates:

    Guidance for populating GenomeTrakr metadata templates (BioSample and SRA)

    9.

    Submit sequence and metadata to NCBI:

    NCBI submission protocol for microbial pathogen surveillance

    10.

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

    NCBI data curation protocol - SOP for editing GenomeTrakr submissions (protocols.io)

    Dry lab - PulseNet submission

    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:

    Populating NCBI template for submissions using BioNumerics v7.6

    14.

    Submit sequence and metadata to NCBI:

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

    15.

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

    NCBI data curation protocol - SOP for editing GenomeTrakr submissions (protocols.io)

    推荐阅读

    Nature Protocols
    Protocols IO
    Current Protocols
    扫码咨询