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URL: https://dx.doi.org/10.17504/protocols.io.bi8dkhs6

Authors: Julie Haendiges, Ruth Timme, Padmini Ramachandran, Maria Balkey

Group: GenomeTrakr

Summary: This procedure outlines the protocol for quantitation of gDNA for subsequent WGS. This document applies to all laboratory personnel in the Division of Microbiology (DM) as well as laboratories in the GenomeTrakr Network.Complete in order:1. DNA Extraction (Manual DNA Extraction or Automated DNA Extraction using the Qiacube)Step-by-step procedures to obtain high quality DNA from isolates in TSB for whole genome sequencing2. DNA Quantitation (included SOP)Quantitation of extracted DNA using the Qubit Flourometer 3. Library Preparation for WGS (Library preparation using Illumina DNA Prep or Library Preparation using Illumina Nextera XT )Library preparation using NexteraXT or Illumina DNA Prep (previously Nextera DNA Flex)2. DNA Quantitation (included SOP)Quantitation of extracted DNA using the Qubit Flourometer 3. Library Preparation for WGS (Library preparation using Illumina DNA Prep or Library Preparation using Illumina Nextera XT )Library preparation using NexteraXT or Illumina DNA Prep (previously Nextera DNA Flex)3. Library Preparation for WGS (Library preparation using Illumina DNA Prep or Library Preparation using Illumina Nextera XT )4. Sequencing using Illumina MiSeq5. Data Quality Checks and NCBI Submission

Affiliations: US Food and Drug Administration, US Food and Drug Administration, US Food and Drug Administration, US Food and Drug Administration

Version: 1

Publication Date: 2020

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