Thermo Fisher Scientific Ion 550 Kit − Chef User guide

Type
User guide
For Research Use Only. Not for use in diagnostic procedures.
Ion 550 Kit − Chef
USER GUIDE
Instructions for automated template preparation, chip loading,
and sequencing
for use with:
IonChef System
Ion S5 XL System
Ion GeneStudio S5 Plus System
Ion GeneStudio S5 Prime System
Catalog NumberA34541
Publication NumberMAN0017275
Revision D.0
Life Technologies Corporation | 5781 Van Allen Way | Carlsbad, California 92008 USA
For descriptions of symbols on product labels or product documents, go to thermofisher.com/symbols-definition.
Revision history: MAN0017275 D.0 (English)
Revision Date Description
D.0 31 July 2023 Updated Ion S5 Chef Supplies (Part No. A27755) kit to include the PCR Plate Frame.
Updated instructions to include loading the PCR Plate Frame. See “Load the pipette tip racks and PCR
Plate” on page31.
Updated instructions to include removing the PCR Plate Frame. See “Remove used consumables” on
page62.
C.0 25 June 2019 Updated for Torrent Suite Software 5.12.
Added #unique_14/unique_14_Connect_42_GUID-440FE1AB-8075-4344-BAD8-69B208711F60 on
page8.
In #unique_45/unique_45_Connect_42_GUID-0796DC72-2287-4303-9088-69174E8FD149, added
clarification for setting up a 1X used Reagents cartridge for a singlechip run 2.
Updated #unique_46/unique_46_Connect_42_GUID-9341033B-85D6-4411-A7D1-B0C72946C8EE to
include the Run Options screen for stepbystep IonChef run setup.
Updated the guidance for storing loaded chips before sequencing in #unique_28/
unique_28_Connect_42_GUID-60C0B2D9-B171-4895-8AE0-D7204463C856.
Added a note in #unique_46/unique_46_Connect_42_GUID-9341033B-85D6-4411-A7D1-B0C72946C8EE
and #unique_28/unique_28_Connect_42_GUID-60C0B2D9-B171-4895-8AE0-D7204463C856 to NOT
remove any residual liquid from chip wells after the IonChef run.
Removed IonChef System maintenance procedures from #unique_97/
unique_97_Connect_42_GUID-47C7590F-FFCD-472A-A61D-9161F491642E.
Updated #unique_97/unique_97_Connect_42_GUID-47C7590F-FFCD-472A-A61D-9161F491642E to add
references to the IonChef System maintenance procedures in IonChef Instrument User Guide.
Updated #unique_120/unique_120_Connect_42_GUID-A2E734CE-DFB9-497D-8BFB-E2FAD0EDF618 to
include the IonChef Instrument User Guide.
B.0 6 December 2018 Updated to include use with the Ion S5 XL System Ion GeneStudio S5 Systems.
Clarification made to dilute barcoded library pools, if used.
Updated for Torrent Suite Software 5.10; Planned Run creation topics reorganized for ease of use.
Ion S5 Controls Kit replaced with Ion S5 Controls Kit Plus.
Removed reference to quality control of ISPs using the Attune NxT Acoustic Focusing Cytometer.
A.0 8 January 2018 New user guide for use with the Ion 550 Kit − Chef, the IonChef System, and the Ion GeneStudio S5 Plus or
Ion GeneStudio S5 Prime System.
The information in this guide is subject to change without notice.
DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE
LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR
ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
Important Licensing Information: This product may be covered by one or more Limited Use Label Licenses. By use of this product,
you accept the terms and conditions of all applicable Limited Use Label Licenses.
Trademarks: All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Guava is a
trademark of Merck KGaA. Agencourt and AMPure are trademarks of Beckman Coulter, Inc. Agilent and Bioanalyzer are trademarks of
Agilent Technologies, Inc. Kimwipes is a trademark of Kimberly-Clark Corporation.
©2018-2023 Thermo Fisher Scientific Inc. All rights reserved.
Contents
CHAPTER1Productinformation .................................................. 7
Productdescription ............................................................. 7
Librarycompatibility ......................................................... 8
Sequencercompatibility ..................................................... 8
Softwarecompatibility ....................................................... 8
Kit contents and storage ......................................................... 8
Kit summary ............................................................... 9
IonChef reagents and materials ............................................. 9
Ion S5 sequencing reagents and materials .................................... 9
Related products .............................................................. 10
Ion 550 Single Chip SupplementalKit ....................................... 10
Compatible Ion Chipkits .................................................. 10
Ion S5 Controls KitPlus ................................................... 10
Required materials notsupplied ................................................. 11
About the Ion Chef System ..................................................... 12
IonChef Instrumentcomponents ........................................... 12
Interior hardware andconsumables .......................................... 13
About the Ion Chef System touchscreen interface ................................. 14
Workflow ..................................................................... 15
CHAPTER2Before youbegin .................................................... 16
Precautions ................................................................... 16
Avoid nucleic acidcontamination ............................................ 16
Avoid chipdamage ........................................................ 16
Protection byequipment .................................................... 16
Types of IonChef runs possible with the Ion 550 Kit −Chef ....................... 17
Guidelines for Ion 550 Kit − Chef reagentuse .................................... 17
Special guidelines for using the Ion 550 Kit − Chef with the IonChefInstrument ..... 18
Prepare the IonChef Instrument foruse ......................................... 18
CHAPTER3Create a PlannedRun .............................................. 19
About PlannedRuns ........................................................... 19
Create a PlannedRun .......................................................... 20
Planned Run workflow keyfields ................................................. 24
Ion 550 Kit − Chef User Guide 3
CHAPTER4Start the IonChefrun ............................................. 26
Materials required .............................................................. 26
Dilute the libraries .............................................................. 26
Guidelines for librarydilution ................................................ 27
Dilute the libraries .......................................................... 27
Prepare theconsumables ....................................................... 28
Add the diluted library to the Library Sample Tube .................................. 29
Load the Ion Chef System ..................................................... 30
Load the pipette tip racks and PCR Plate ..................................... 31
Load the Reagents cartridges—singlechiprun ................................ 33
Load the Reagents cartridge—dualchiprun .................................. 34
Load the Solutions cartridge ................................................ 36
Load the Recovery Tubes and Enrichment Cartridgev2 ......................... 36
Load the Chip-loading centrifuge—singlechiprun ............................. 38
Load the Chip-loading centrifuge—dualchiprun .............................. 39
Confirm that consumables are correctlyinstalled .............................. 42
IonChef pre-runchecklist ................................................. 43
Start the IonChefrun ......................................................... 44
Unload the chips forsequencing ................................................. 50
CHAPTER5Initialize thesequencer ............................................. 52
Ion GeneStudio S5 System componentpositions ................................. 52
Before you begin .............................................................. 53
When a manual cleaning of the sequencer is required .............................. 53
Initialize thesequencer ......................................................... 54
CHAPTER6Start the sequencingrun ........................................... 56
Chip handlingguidelines ........................................................ 56
Start the sequencing run ....................................................... 57
Maintain thesequencer ......................................................... 59
Materials required .......................................................... 59
Clean or decontaminate thesequencer ....................................... 59
Reagent consumablesdisposal .................................................. 59
CO2 scrubber removal anddisposal .......................................... 59
Recycle Ion S5 Wash Solution and Ion S5 Cleaning Solutionbottles ........... 60
Contents
4Ion 550 Kit − Chef User Guide
CHAPTER7Clean the IonChef System ...................................... 61
About the cleaning protocol ..................................................... 61
Materials required .............................................................. 61
Clean the IonChefInstrument .................................................. 61
Remove usedconsumables ................................................. 62
Inspect and clean the Recovery centrifuges and buckets ....................... 64
Start the cleaning ......................................................... 65
APPENDIXATroubleshooting .................................................... 67
View troubleshooting and FAQsonline ............................................ 67
Ion Chef System setup and operation ........................................... 67
Ion GeneStudio S5 System setup and operation .................................. 70
Instrument alarms andevents ............................................... 70
Initialization—General errors ................................................ 70
Ion Sphere Assay troubleshootingtable ......................................... 71
APPENDIXBSupplemental procedures ........................................ 72
Perform a manual cleaning of thesequencer ...................................... 72
Perform an instrument reset run with an initialized sequencer that is loaded with
an unused Reagents cartridge ................................................. 73
Quality control of Ion 550 ISPs ................................................. 75
Quality control using the Ion Sphere Assay on the Qubit Fluorometer .......... 75
Acceptance criteria for unenriched Ion 550 ISPs ............................. 75
Quality control using the Guava easyCyte 5 Benchtop Flow Cytometer .......... 75
Maintain the IonChef System .................................................. 77
APPENDIXCSafety ............................................................... 78
Symbols on thisinstrument ..................................................... 78
Conformitysymbols ........................................................ 80
Location of safety labels on thisinstrument ........................................ 80
Safety information for instruments not manufactured by Thermo FisherScientific ...... 80
Instrumentsafety .............................................................. 80
General ................................................................... 80
Physical injury ............................................................. 81
Electricalsafety ........................................................... 81
Cleaning anddecontamination .............................................. 82
Ultraviolet (UV)Safety ...................................................... 82
Safety and electromagnetic compatibility (EMC) standards .......................... 82
Safetycompliance ......................................................... 82
EMC ..................................................................... 83
Environmentaldesign ...................................................... 83
Contents
Ion 550 Kit − Chef User Guide 5
Instrument safety — Ion GeneStudio S5 System ................................. 83
Chemicalsafety ................................................................ 84
Biological hazardsafety ......................................................... 85
Documentation and support ....................................................... 86
Relateddocumentation ......................................................... 86
Customer and technical support ................................................. 87
Limited product warranty ........................................................ 87
Contents
6Ion 550 Kit − Chef User Guide
Product information
Productdescription .................................................................... 7
Kit contents and storage ............................................................... 8
Related products ..................................................................... 10
Required materials notsupplied ........................................................ 11
About the Ion Chef System ........................................................... 12
About the Ion Chef System touchscreen interface ....................................... 14
Workflow ............................................................................ 15
IMPORTANT! Before using this product, read and understand the information in the “Safety” appendix
in this document.
Product description
The Ion 550 Kit − Chef (Cat. No. A34541) contains the reagents and materials that are required to
perform the following steps in the sequencing workflow.
Prepare enriched, template-positive Ion Sphere Particles (ISPs) and load them onto an Ion 550
Chip using the IonChef Instrument.
Sequence the loaded chips on the Ion GeneStudio S5 Plus Sequencer, Ion GeneStudio S5
Prime Sequencer, or Ion S5XL Sequencer.
IMPORTANT! The Ion 550 Kit − Chef and Ion 550 Chips are not compatible with the Ion S5
System or the base model Ion GeneStudio S5 System.
Each kit contains all the supplies that are required to load and sequence 8 chips and is compatible for
use with up to 200-base-read libraries. The kit supports 4 IonChef runs (2 chips/IonChef run) and 4
sequencer initializations (2 sequencer runs/sequencer initialization).
To provide more workflow flexibility, the Ion 550 Chef Reagents and Ion 550 Chef Solutions
cartridges in the Ion 550 Kit − Chef are now configured for both standard dualchip templating runs,
and single-chip templating runs. The Ion 550 Chef Reagents and Ion 550 Chef Solutions cartridges
can be reused in a second single-chip templating run if the second run is performed within 8days of
the first run.
The Ion 550 Single Chip Supplemental Kit must be ordered separately to obtain the additional single-
use supplies that are required for 8singlechip templating runs and 8 sequencer initializations. For
ordering information for this kit, for the Ion 550 Chip Kit, and for the Ion S5 Controls Kit Plus, see
“Related products” on page10.
1
Ion 550 Kit − Chef User Guide 7
Library compatibility
The Ion 550 Kit − Chef can be used with up to 200-base-read libraries of any type prepared using any
available Ion Torrent-branded library kit.
Sequencer compatibility
The Ion 550 Chips are compatible with the Ion S5 XL System, Ion GeneStudio S5 Plus System, and
Ion GeneStudio S5 Prime System. The Ion 550 Chips are NOT compatible with the Ion S5 System
or the Ion GeneStudio S5 System.
Software compatibility
The Ion 550 Kit − Chef is compatible with Torrent Suite Software 5.8 and later. Be sure to update
your Torrent Suite Software to the latest available version before using this kit. For more information,
see the Torrent Suite Software User Guide for your version of the software.
Kit contents and storage
IMPORTANT! Do not substitute components with components from other Ion sequencing kits. We
have verified this protocol using these specific materials. Substitution can adversely aect system
performance.
IMPORTANT! Store all consumables and cartridges under the recommended conditions and in an
upright position. Do NOT store the Ion S5 Sequencing Reagents (Cat.No.A27768) on dry ice or in a
closed environment where dry ice is present.
Each Ion 550 Kit − Chef contains all the supplies that are required to prepare and sequence 8 Ion
550 Chips. Cat. No. A34541 supports 4 IonChef runs (2 chips/IonChef run) and 4 sequencer
initializations (2 sequencing runs/sequencer initialization) for up to 200base-read libraries.
To support 8 single-chip IonChef runs and 8 sequencer initializations with the Ion 550 Kit − Chef,
order the Ion 550 Single Chip Supplemental Kit (see page10).
On arrival, inspect all consumables. Contact Technical Support if any of the products have been
damaged during shipping.
Chapter1Product information
Kit contents and storage
1
8Ion 550 Kit − Chef User Guide
Kit summary
Component PartNo. Quantity per kit
Ion S5 Chef Supplies A27755 4 boxes
Ion 550 Chef Solutions A36410 1 box
Ion 550 Chef Reagents A34540 1 box
Ion S5 Sequencing Solutions A27767 1 box
Ion S5 Sequencing Reagents A27768 1 box
IonChef reagents and materials
Contents Amount / box Storage
Ion S5 Chef Supplies (Part No. A27755)
Chip Adapter 2 15°C to 30°C
Enrichment Cartridge v2 1
Tip Cartridge v2 1
PCR Plate 1
PCR Plate Frame 1
Frame Seal v2 1
Recovery Station Disposable Lid v2 2
Recovery Tube v2 12
Ion 550 Chef Solutions (Part No. A36410)
Ion 550 Chef Solutions 4 cartridges 15°C to 30°C
Ion 550 Chef Reagents (Part No. A34540)
Ion 550 Chef Reagents 4 cartridges –30°C to –10°C
Ion S5 sequencing reagents and materials
Contents Amount / box Storage
Ion S5 Sequencing Solutions (Part No. A27767)
Ion S5 Wash Solution 4×1.5L 15°C to 30°C
Ion S5 Cleaning Solution 250mL
Chapter1Product information
Kit contents and storage 1
Ion 550 Kit − Chef User Guide 9
Contents Amount / box Storage
Ion S5 Sequencing Reagents (Part No. A27768)
Ion S5 Sequencing Reagents 4 cartridges –30°C to –10°C[1]
[1] Cartridges ship at 2°C to 8°C. Store as indicated, donot store on dry ice.
IMPORTANT! Do not store the Ion S5 Sequencing Reagents (PartNo.A27768) on dry ice or in a
closed environment containing dry ice.
Related products
Ion 550 Single Chip Supplemental Kit
The Ion 550 Single Chip Supplemental Kit (Cat. No. A36953), when used with the Ion 550 Kit − Chef,
provides reagents and materials for eight single-chip IonChef runs and eight sequencer initializations.
The Ion 550 Single Chip Supplemental Kit is ordered separately.
Contents Part No. Quantity per kit Storage
Ion S5 Chef Supplies A27755 4 boxes 15°C to 30°C
Ion S5 Sequencing Solutions A27767 1 box
Ion S5 Sequencing Reagents A27768 1 box −30°C to −10°C
Compatible Ion Chip kits
Description Cat. No. Quantity per kit Storage
Ion 550 Chip Kit (4-pack) A34537 4 chips 15°C to 30°C
Ion 550 Chip Kit (2×4-pack) A34538 8 chips
Ion S5 Controls Kit Plus
The Ion S5 Controls Kit Plus (Cat. No. A30729) is ordered separately. See the Ion S5 Controls Kit Plus
Product Information Sheet (Pub. No. MAN0016206) for details.
Contents Amount Storage
Human CEPH Genomic DNA Control (red cap) 30 µL −30°C to −10°C
Human CEPH Control 200 Library (yellow cap) 12 µL
E. coli DH10B Control 400 Library (orange cap) 16 µL
Chapter1Product information
Related products
1
10 Ion 550 Kit − Chef User Guide
Required materials not supplied
Unless otherwise indicated, all materials are available through thermofisher.com. "MLS" indicates that
the material is available from fisherscientific.com or another major laboratory supplier.
Item Source
IonChef System 4484177
Microcentrifuge[1] MLS
2, 20, 200, and 1,000µL pipettes and appropriate filtered tips MLS
Microcentrifuge tubes, 1.5-mL or 1.7-mL MLS
Vortex mixer with a rubber platform MLS
Gloves, powder-free nitrile MLS
Ice buckets and ice
Nuclease-free water, molecular biology grade MLS
Isopropyl alcohol, 70% solution MLS
Wipes, disposable lint-free MLS
(Optional) Uninterruptible Power Supply (UPS) [2] MLS
[1] Must fit standard 0.2- and 1.5mL microcentrifuge tubes and generate 15,500 × g. To convert the RPMs of your centrifuge to RCF in
units of gravity, see tools.thermofisher.com/content/sfs/brochures/TR0040-Centrifuge-speed.pdf.
[2] For laboratories that experience frequent power outages or line voltage fluctuations, we recommend that you use an uninterruptible
power supply that is compatible with 2500 W output or higher.
Chapter1Product information
Required materials not supplied 1
Ion 550 Kit − Chef User Guide 11
About the Ion Chef System
The Ion Chef System (Cat. No. 4484177) provides automated, high-throughput template preparation
and chip loading for use with the Ion S5XL Sequencer, Ion GeneStudio S5 Plus Sequencer, or
Ion GeneStudio S5 Prime Sequencer. The system includes a set of cartridge-based consumables
and reagents that enable a user to load two Ion 550 Chips in approximately 16hours with less than
15 minutes of hands-on time. The Ion Chef System features network integration with the Torrent
Suite Software to enable sample and reagent traceability throughout the chip preparation workflow.
IonChef Instrument components
The following figure illustrates the major external and internal features of the IonChef Instrument.
4
1 2 3
5 6 7
8
1Door – Provides access to the interior of the
instrument. The door is locked in the closed position
during operation.
2Micropipettor – A mechanical positive-displacement
pipettor that performs all fluid transfers during sample
and chip preparation.
3Robotic arm – Enables fluid transfer by the
Micropipettor. The arm also contains an optical sensor
that reads the barcodes of instrument reagents and
consumables.
4Touchscreen – Provides access to all instrument
functions for operation, maintenance, and
troubleshooting.
5Power button – Power switch for the IonChef
Instrument, where the states are on (illuminated) and
o.
6Power port – A 100–240 VAC port that provides power
to the IonChef Instrument.
7Ethernet port – An RJ45 port that provides Ethernet
(100Mbit) communication with the IonChef
Instrument.
8USB port – Provides USB communication with the
IonChef Instrument. Used to update the instrument
firmware and to transfer data during service or
maintenance.
Chapter1Product information
About the Ion Chef System
1
12 Ion 550 Kit − Chef User Guide
Interior hardware and consumables
The following figure illustrates the interior of the IonChef Instrument and describes the stations
involved in the preparation of chips for sequencing on an Ion S5 XL System, Ion GeneStudio S5 Plus
System, or Ion GeneStudio S5 Prime System.
A
B
C
D
E
F
G
H
1 2 3 4 56 7 8 9 10 11 12
5
3
8
12
6
4
7
530 CCBD02226
530 CCBD02226
9
1Waste pipette tip rack – The position of the rack containing waste (used) pipette tips.
2Automated heated cover – Transfers the plate cover to the PCR reaction plate within the sample block. During
thermal cycling, the heated cover applies compression to seal the reaction plate and heats the cover to prevent
condensation.
3New pipette tips – The position of the rack containing unused pipette tips.
4Thermal cycler sample block – Performs thermal cycling of the sequencing reactions on a 96-well PCR reaction
plate.
5Reagents station – The position on the instrument deck of the diluted libraries, NaOH, and the Ion 550 Chef
Reagents cartridge.
6Solutions station – The position on the instrument deck of the Ion 550 Chef Solutions cartridge, which is
maintained at room temperature.
7Recovery centrifuges – Twin stations that perform centrifugation of the ISPs during the recovery phase of template
preparation.
8Enrichment station – The position of the rack containing consumables for enrichment of the template-positive ISPs.
9Chip-loading centrifuge – Performs centrifugation of sequencing chips that have been mounted to chip-loading
adapters and loaded with template-positive ISPs.
Chapter1Product information
About the Ion Chef System 1
Ion 550 Kit − Chef User Guide 13
About the Ion Chef System touchscreen interface
The Ion Chef System features a simple interface for loading chips, cleaning the instrument, and
performing system maintenance and configuration tasks.
2
1
3 4 5
1Set up run button – Set up the IonChef template preparation and chip-loading routine. Choose Step by Step to
have the instrument lead you stepwise through installation of reagents and consumables, or choose Quick Start to
proceed if you have already installed the consumables.
2Open Door button.
3Notifications button – View notifications about instrument status during and between runs.
4Quick Start button – Proceed directly to the Quick Start instrument setup mode. User verifies the loading of a new
pipette tip cartridge and an empty pipette tip rack to hold waste tips generated during the run, before proceeding to
Deck Scan.
5Settings button – Advance to a screen providing the following options:
Notifications: view notifications about instrument status during and between runs
Instrument Settings: view current settings and network configuration, set instrument name, adjust time zone
Service tools: access screens for service-related maintenance and instrument diagnostics
Torrent Server: add and manage Ion Torrent Server connections
Clean Ion Chef: proceed directly to the instrument cleaning routine
Check for updates: check availability of system software updates
Chapter1Product information
About the Ion Chef System touchscreen interface
1
14 Ion 550 Kit − Chef User Guide
Workflow
The following workflow shows how to prepare and load samples using the Ion 550 Kit − Chef and Ion
Chef System for sequencing on an Ion S5XL Sequencer, Ion GeneStudio S5 Plus Sequencer, or
Ion GeneStudio S5 Prime Sequencer.
Ion 510 & Ion 520 & Ion 530 Kit – Chef Workflow
Create a Planned Run (page19)
Create a planned run in Torrent Suite Software by entering run
parameters for templating and sequencing your samples.
Dilute the library samples (page26)
If needed, dilute sample and control libraries. Thaw the Reagents
cartridge at room temperature for 45 minutes before use.
Load the IonChef Instrument (page30)
Load the IonChef Instrument with sequencing chips, reagents, and
sample libraries.
Start the IonChef run (page26)
Start the IonChef run by following the step-by-step instructions on the
IonChef Instrument home touchscreen.
Unload the IonChef Instrument (page50)
After the IonChef Run completes, unload the prepared sequencing
chips from the IonChef Instrument.
Load the prepared sequencing chip into the
sequencer and begin the sequencing run
(page56)
Load the prepared sequencing chips on a sequencer and follow the
instructions on the touchscreen to start the sequencing run.
Chapter1Product information
Workflow 1
Ion 550 Kit − Chef User Guide 15
Before you begin
Precautions .......................................................................... 16
Types of IonChef runs possible with the Ion 550 Kit −Chef ............................ 17
Guidelines for Ion 550 Kit − Chef reagentuse .......................................... 17
Special guidelines for using the Ion 550 Kit − Chef with the IonChefInstrument .......... 18
Prepare the IonChef Instrument foruse ............................................... 18
Precautions
Avoid nucleic acid contamination
IMPORTANT! A primary source of contamination is spurious DNA fragments from previous sample
processing steps. Do not introduce amplified DNA into the library preparation laboratory or work area.
Avoid chip damage
IMPORTANT! To avoid possible damage to the chip due to electrostatic discharge, ground yourself
before picking up a chip or placing a chip on a surface such as a lab bench. For example, touch the
metal trim on the chip compartment before inserting or removing a chip from the chip clamp.
Protection by equipment
WARNING! The protection that is provided by the equipment can be impaired if the instrument is
operated outside the environment and use specifications, the user provides inadequate maintenance,
or the equipment is used in a manner that is not specified by the manufacturer (Thermo Fisher
Scientific).
2
16 Ion 550 Kit − Chef User Guide
Types of IonChef runs possible with the Ion 550 Kit −
Chef
Three types of IonChef templating runs can be performed using the Ion 550 Kit − Chef, depending
on the number of chips that are loaded, and whether a single-chip run is the first or second run with
a set of Reagents and Solutions cartridges. The IonChef Instrument tracks cartridge usage and uses
the following names for the types of runs that are allowed with a given set of cartridges:
Single-chip run 1—a run loading a single chip using a new set of cartridges
Single-chip run 2—a run loading a single chip using a set of cartridges that have already been used
in a single-chip run 1
Dual-chip run—a run loading two chips using a new set of cartridges
Guidelines for Ion 550 Kit − Chef reagent use
To ensure the proper function of the IonChef System, handle the associated consumables and
reagents according to the following guidelines:
Store all IonChef System consumables and cartridges under the recommended conditions and in
an upright position.
Inspect all IonChef System consumables and cartridges for damage on arrival and again before
use.
Hold sequencing chips by gently gripping them by their edges.
When the IonChef System is not in use, remove all consumables and reagents from the deck and
close the instrument door.
After performing a single-chip run 1, store the Ion 550 Chef Reagents cartridge used in the run
at 4°C. Store the Ion 550 Chef Solutions cartridge used in the run at 15°C to 30°C. Use the
cartridges in a single-chip run 2 within eight days of the first run.
If you thaw a Ion 550 Chef Reagents cartridge, and then decide not to perform an IonChef run,
do not return the cartridge to the freezer for storage at −30°C to −10°C. Store a thawed Reagents
cartridge at 4°C. Use the cartridge for a dualchip run, or for both single-chip runs within eight days
of thawing.
It is good practice to label Reagents and Solutions cartridges that have been used in a single-chip
run 1 with the date of the run.
When performing a dual-chip run, always load two Ion 550 Chips in the IonChef Instrument. You
cannot load an Ion 550 Chip with another chip type in an IonChef run.
Use only Ion Torrent kits and supplies with the IonChef Instrument. The use of third-party
reagents and supplies can adversely aect the performance of the IonChef Instrument and chips
prepared.
Chapter2Before you begin
Types of IonChef runs possible with the Ion 550 Kit − Chef 2
Ion 550 Kit − Chef User Guide 17
Remove and sequence chips as soon as possible after the IonChef System finishes loading
them. If you cannot sequence a loaded chip immediately, store it in a chip storage container at 4°C
until you are ready to run it (up to 24hours maximum).
IMPORTANT! If you choose to store a loaded chip, remove the chip from 4°C storage (but keep
it in the chip storage container) at least 20 minutes before running it, allowing the chip to warm to
room temperature.
If you sequence a second chip more than 24 hours after sequencer initialization was performed for
the first chip, you must reinitialize the sequencer.
Special guidelines for using the Ion 550 Kit − Chef with the
IonChef Instrument
IonChef Instrument users must create a Planned Run for each chip in a templating run. Planned
Runs are not optional for IonChef runs with the Ion 550 Chip.
The IonChef Instrument must be connected to a Ion Torrent Server to enable cartridge use
tracking and proper use of the single-chip workflow with the Ion 550 Kit − Chef.
Note: If the instrument is not connected, see IonChef Instrument Network Connection User
Guide (Pub. No. MAN0013444) for instructions on how to configure a direct or indirect network
connection of the IonChef Instrument to a server.
To use the flexible workflow feature and enable cartridge use tracking between Ion Torrent
Servers across multiple IonChef Instruments, servers must be linked in an Ion Mesh
configuration. For further information on how to link servers in an Ion Mesh setup, go to Software
Administration4Set up Ion Mesh in Torrent Suite Software Help.
IMPORTANT! If you have servers connected in an Ion Mesh configuration, and the connection
to one server is disrupted or lost, cartridge tracking between servers is disabled. In this situation,
an error message appears if you attempt to start a run. The run is not allowed to start because
cartridge use status is not trackable. Ion Mesh communication must be restored to verify cartridge
use status before the run can start.
Prepare the IonChef Instrument for use
Ensure that the IonChef Instrument has been cleaned following the previous run. If not, clean the
instrument before loading it with consumables.
Note: For more information on the cleaning procedure, see Chapter7, “Clean the IonChef
System”.
Inspect the empty compartments of the Reagents and Solutions stations for condensation.
Condensate can collect in these compartments, depending on temperature and humidity
conditions. Before loading consumables into the instrument, wipe the compartments dry with a
laboratory wipe or absorbent cloth, if needed.
Thaw the Ion 550 Chef Reagents cartridge at room temperature for 45 minutes before use.
Chapter2Before you begin
Special guidelines for using the Ion 550 Kit − Chef with the IonChef Instrument
2
18 Ion 550 Kit − Chef User Guide
Create a Planned Run
About PlannedRuns .................................................................. 19
Create a PlannedRun ................................................................. 20
Planned Run workflow keyfields ....................................................... 24
About Planned Runs
Planned Runs are digital instructions that are created in Torrent Suite Software for controlling the
template preparation and sequencing instruments. Planned Runs contain settings such as number
of flows, kit types, barcodes, sample information, and reference files (if any). Planned Runs are also
used to track samples, chips, and reagents throughout the workflow, from template preparation on
the Ion Chef Instrument through sequencing on an Ion S5XL Sequencer, Ion GeneStudio S5 Plus
Sequencer, or Ion GeneStudio S5 Prime Sequencer and subsequent data analysis. Each chip that is
prepared in an IonChef run requires its own Planned Run.
IMPORTANT! For more information on creating a Planned Run in Torrent Suite Software, including
a complete description of each field in the Create Plan workflow bar, see the Torrent Suite Software
Help, available by clicking the Help button in the software.
3
Ion 550 Kit − Chef User Guide 19
Create a Planned Run
IMPORTANT! If you are using the Ion 550 Kit − Chef with libraries that were prepared
using Oncomine panels, see the corresponding Oncomine Assay User Guide for assay-specific
instructions on creating a Planned Run in Torrent Suite Software.
1. Sign in to the Torrent Suite Software.
2. In the Plan tab, in the Templates screen, select the application that you want to run (such as
AmpliSeq DNA) from the left navigation menu, then click one of the following options:
Plan New Run—to plan a new run using the Generic Sequencing template for the selected
application.
(Actions)4Plan Run in the row of the template—to plan a new run using a specific
Planned Run template.
3. In the Create Plan workflow bar, review the IonReporter and Research Application steps, then
make selections appropriate to your run. Click Next.
4. In the Kits step, make the following selections:
a. Select Ion GeneStudio S5 System from the Instrument dropdown list.
b. Select the Ion 550 Chip from the Chip Type dropdown list.
c. Select the library kit used to prepare your libraries from the Library Kit Type dropdown list.
d. (Optional) For barcoded libraries, select the barcode set used during library preparation from
the Barcode Set dropdown list.
Chapter3Create a Planned Run
Create a Planned Run
3
20 Ion 550 Kit − Chef User Guide
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Thermo Fisher Scientific Ion 550 Kit − Chef User guide

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