Aug 23, 2024

Public workspaceSEQUENCING Protocol Template V.3

This protocol is a draft, published without a DOI.
  • 1MBARI;
  • 2AWI
  • Better Biomolecular Ocean Practices (BeBOP)
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Protocol CitationKathleen Pitz, Raïssa Meyer 2024. SEQUENCING Protocol Template. Protocol exchange https://protocols.io/view/sequencing-protocol-template-dj234qgnVersion created by Kathleen Pitz
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: November 14, 2023
Last Modified: August 23, 2024
Protocol Integer ID: 106299
Abstract
A protocol template created through the BeBOP project for sequencing.

Main protocol steps begin below the "Standard Operating Procedure" section. A detailed description of the BeBOP project and these templates is available here: https://github.com/BeBOP-OBON/
MIOP: Minimum Information about an Omics Protocol
MIOP: Minimum Information about an Omics Protocol

MIOP TermValue
methodology category
project
purpose
analyses
geographic location
broad-scale environmental context
local environmental context
environmental medium
target
creator
materials required
skills required
time required
personnel required
language
issued
audience
publisher
hasVersion
license
maturity level
See MIOP_definition.md for term definitions. The terms "methodology category" and "maturity level" have controlled vocabulary.

AUTHORS
AUTHORS

PREPARED BY All authors known to have contributed to the preparation of this protocol, including those who filled in the template.AFFILIATIONORCID (visit https://orcid.org/ to register)DATE
Content CellContent CellContent Cellyyyy-mm-dd
Content CellContent CellContent Cellyyyy-mm-dd

RELATED PROTOCOLS
RELATED PROTOCOLS

PROTOCOL NAME AND LINKISSUER / AUTHORRELEASE / ACCESS DATE
Content CellContent Cellyyyy-mm-dd
Content CellContent Cellyyyy-mm-dd
This is a list of other protocols which should be known to users of this protocol. Please include the link to each related protocol.

ACRONYMS AND ABBREVIATIONS
ACRONYMS AND ABBREVIATIONS

ACRONYM / ABBREVIATIONDEFINITION
Content CellContent Cell

GLOSSARY
GLOSSARY

SPECIALISED TERMDEFINITION
Content CellContent Cell
Content CellContent Cell

BACKGROUND
BACKGROUND
Summary
Insert a short description of the background for the method/protocol (e.g. why and for which purpose do you perform sequencing). Please provide a brief summary of your method including, as appropriate, a brief description of what techniques your best practice is about, which ocean environments or regions it targets, the primary sensors covered, what type of data/measurements/observing platform it covers, limits to its applicability.
Method description and rationale
Insert a short description of the functioning principal of the methodology used in the protocol (i.e. how does the method work?). Please note that this is different from the step-by-step description of the protocol procedure. Insert a short statement explaining why the specific methodology used in the protocol has been selected (e.g. it is highly reproducible, highly accurate, procedures are easy to execute etc….).
Spatial coverage and environment(s) of relevance
If applicable, please specify the region where the protocol is applied. For regional term guidance see here. If applicable, please indicate here the environment(s) of relevance for the protocol, e.g. Abyssal plain. Select from the ENVO terminology.
PERSONNEL REQUIRED
Insert the number of technicians, data managers, and scientists required for the good execution of the procedure
Safety
Identify hazards associated with the procedure and specify protective equipment and safety training required to safely execute the procedure
Training requirements
Specify technical training required for the good execution of the procedure.
Time needed to execute the procedure
Specify how much time is necessary to execute the procedure.
EQUIPMENT
EQUIPMENT

DESCRIPTION e.g. filterPRODUCT NAME AND MODEL Provide the official name of the productMANUFACTURER Provide the name of the manufacturer of the product.QUANTITY Provide quantities necessary for one application of the standard operating procedure (e.g. number of filters).REMARK For example, some of the consumable may need to be sterilized, some commercial solution may need to be diluted or shielded from light during the operating procedure.
Durable equipment
Content CellContent CellContent CellContent CellContent Cell
Content CellContent CellContent CellContent CellContent Cell
Consumable equipment
Content CellContent CellContent CellContent CellContent Cell
Content CellContent CellContent CellContent CellContent Cell
Chemicals
Content CellContent CellContent CellContent CellContent Cell
Content CellContent CellContent CellContent CellContent Cell

STANDARD OPERATING PROCEDURE
STANDARD OPERATING PROCEDURE
In the following SOP, please use the exact names of equipment as noted in the table above.

Provide a step-by-step description of the protocol. The identification of difficult steps in the protocol and the provision of recommendations for the execution of those steps are encouraged.
PREPARATION
PREPARATION
Please specify the preparatory actions you took before you collected the samples and note what equipment was needed to do so (e.g. disinfection of work surfaces, preparations to the equipment you intend to use later on).

Step
Step
Step
SEQUENCING
SEQUENCING
Please specify the actions you took to sequence the DNA and note what equipment was needed to do so (loading of sequencing machine, details about the sequencing run, sequencing institution (if out of house), sequencing primers used).

Step

Sequencing Primer NameDirectionSequence (5’ -> 3’)
contentforwardcontent
contentreversecontent

Step
QUALITY CONTROL
QUALITY CONTROL
Describe and explain criteria used to validate results of the standard operating procedure.
BASIC TROUBLESHOOTING GUIDE
BASIC TROUBLESHOOTING GUIDE
Identify known issues associated with the procedure, if any. Provide troubleshooting guidelines when available.
REFERENCES
REFERENCES
Insert all references cited in the document. Please insert full DOI address when available, e.g. http://doi.dx.org/10.1007/s11258-014-0404-1
APPENDIX A: DATASHEETS
APPENDIX A: DATASHEETS
Link templates (e.g. preformatted spreadsheets) used to record measurements and report on the quality of the data as well as any documents such as manufacturer specifications, images, etc that support this protocol. Please include a short note describing the document’s relevance.