Thermo Fisher Scientific Ion PGM Hi?Q View Sequencing Kit User guide

Brand: Thermo Fisher Scientific

Model: Ion PGM Hi?Q View Sequencing Kit

Type: User guide

For Research Use Only. Not for use in diagnostic procedures.

Ion PGM™ Hi‑Q™ View Sequencing Kit

USER GUIDE

for use with:

Ion PGM™ Hi‑Q™ View Sequencing Kit

Ion PGM™ System

Ion 318™ Chip v2 BC

Ion 316™ Chip v2 BC

Ion 314™ Chip v2 BC

Catalog Number A30044

Publication Number MAN0014583

Revision C.0

The information in this guide is subject to change without notice.

DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, LIFE TECHNOLOGIES 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.

Revision history: Pub. No.MAN0014583

Revision Date Description of change

C.0 6 January 2017 • Information in appendices C−G moved to the

Ion PGM

™

System Reference Guide

(Pub. No.

MAN0009783)

• Updated web links and references

B.0 31 August 2016 • Correction of minor errors in component names and amount provided.

• Recommendation for the time that can elapse between initialization and the start of the ﬁnal

run changed from 27 hours to 24 hours

A.0 8 April 2016 New User Guide, which includes instructions to use the new Ion PGM™ Hi‑Q™ View Sequencing Kit

with the Ion PGM™ System

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.

Corporate entity: Life Technologies Corporation | Carlsbad, CA 92008 USA | Toll Free in USA 1 800 955 6288

Trademarks: All trademarks are the property of Thermo Fisher Scientiﬁc and its subsidiaries unless otherwise speciﬁed. Rainin and Pipet-Lite are

trademarks of Rainin Instrument, LLC. ELGA and PURELAB are trademarks of VWS (UK) Ltd. Luer-Lok is a trademark of Becton, Dickinson and

Company Corporation.

Contents

■CHAPTER 1 Product Information ....................................... 6

Product description ............................................................. 6

Intended use ................................................................... 7

Library kit compatibility .......................................................... 7

Template kit compatibility ........................................................ 7

Software and instrument system compatibility ...................................... 7

Kit contents and storage ......................................................... 8

Ion PGM™ Wash 2 Bottle Kit ....................................................... 9

Ion Chip kits .................................................................... 9

Instruments and server .......................................................... 9

Required materials not supplied ................................................. 10

Ion PGM™ System layout ........................................................ 12

■CHAPTER 2 Before you begin .......................................... 13

Before you begin ............................................................... 13

Update the software ........................................................ 13

Instrument installation by trained personnel only .............................. 13

Nucleic acid contamination .................................................. 13

CO2 contamination ......................................................... 13

Instrument vibration and clearances ......................................... 13

Static electricity ........................................................... 14

Ventilation requirements ................................................... 14

Guidelines for sequencing runs .................................................. 14

Guidelines to prevent cross-contamination ........................................ 14

Guidelines for pipetting ......................................................... 15

Guidelines for chip handling and use ............................................. 15

Gas cylinders .................................................................. 15

Perform a leak test ........................................................ 16

Protocol workﬂow .............................................................. 17

■CHAPTER 3 Create a Planned Run .................................... 18

About Planned Runs ............................................................ 18

Create a Planned Run .......................................................... 19

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

■CHAPTER 4 Clean and initialize ....................................... 21

Condition the Wash 2 Bottle for ﬁrst use .......................................... 21

Clean the Ion PGM™ System ..................................................... 21

Materials required ......................................................... 21

Cleaning schedule ......................................................... 22

Cleaning setup ............................................................ 22

18 MΩ water cleaning ...................................................... 22

Chlorite cleaning .......................................................... 23

Initialize the Ion PGM™ System ................................................... 24

Materials required ......................................................... 24

Initialization guidelines ..................................................... 25

Before initialization ........................................................ 25

Prepare the Wash 2 Bottle .................................................. 25

Prepare the Wash 1 and Wash 3 Bottles ....................................... 27

Begin the initialization ...................................................... 28

Prepare the 50‑mL Reagent Bottles with dNTPs ............................... 30

Attach the sipper tubes and Reagent Bottles .................................. 30

■CHAPTER 5 Load the chip and start the sequencing run ............ 32

Materials required ............................................................. 32

Before starting ................................................................ 32

Add controls to the enriched, template-positive ISPs ............................... 33

Anneal the Sequencing Primer .................................................. 33

Perform Chip Check ............................................................ 34

Bind the Sequencing Polymerase to the ISPs ...................................... 36

Prepare and load the chip ....................................................... 37

Remove liquid from the chip ................................................. 37

Load the chip .............................................................. 38

Select the Planned Run and perform the run ...................................... 41

Select the Planned Run ..................................................... 41

Perform the run ........................................................... 42

■APPENDIX A Troubleshooting ......................................... 43

Chip Check .................................................................... 43

Chip calibration (before loading sample) .......................................... 44

Chip calibration (after loading sample) ............................................ 46

Initialization—General errors .................................................... 47

Initialization—Auto pH errors .................................................... 48

Initialization—Reagent pH veriﬁcation ............................................ 53

Troubleshooting using Control Ion Sphere™ Particles and control libraries ............ 54

Contents

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

■APPENDIX B Sharing Planned Runs between Torrent Servers ..... 55

Enable Planned Run sharing ..................................................... 55

Set up Server Network (admin action) ........................................ 56

Transfer a Planned Run .................................................... 57

Undo a Planned Run transfer (administrator) .................................. 59

Undo a Planned Run transfer (user) .......................................... 59

■APPENDIX C Additional instrument information .................... 60

■APPENDIX D Safety ..................................................... 61

Instrument safety .............................................................. 62

Chemical safety ................................................................ 62

Biological hazard safety ......................................................... 63

■Documentation and support ............................................. 64

Customer and technical support ................................................. 64

Limited product warranty ....................................................... 64

Contents

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Product Information

CAUTION! ABBREVIATED SAFETY ALERTS. Hazard symbols and hazard

types specied in procedures may be abbreviated in this document. For the

complete safety information, see the “Safety” appendix in this document.

IMPORTANT! Before using this product, read and understand the information in the

“Safety” appendix in this document.

Product description

The Ion PGM™ Hi‑Q™ View Sequencing Kit (Cat. No. A30044) includes reagents and

materials for sequencing up to ~400‑bp inserts using the following chips on the Ion

PGM™ System:

• Ion 318™ Chip v2 BC

• Ion 316™ Chip v2 BC

• Ion 314™ Chip v2 BC

The Ion PGM™ Hi‑Q™ View Sequencing Kit is designed for use with the Ion PGM™

Hi‑Q™ View OT2 Kit, which introduces colored Ion PGM™ Hi‑Q™ View ISPs for easier

sample visualization. In addition, the Ion PGM™ Hi‑Q™ View Sequencing Solutions

show improved performance in terms of reduced strand bias in amplicon sequencing.

Note: The kit and chips may also be used for Research Use Only experiments on the

Ion PGM™ Dx System running Torrent Suite™ Assay Development Software[1].

The kit includes components for 4 initializations. The number of sequencing runs per

kit depends on the length of the base reads, as shown in the following table.

Sequencing type Number of ﬂows Number of runs per

kit

Maximum number

of runs per

initialization[1]

500-base-read[2] 1100 4 1

400-base read 850 4 1

200-base read 500 8 2

£100-base read 260 12 3

[1] For best results, begin the run within 1 hour after initialization.

[2] For ISPs prepared using the Ion PGM™ Template IA 500 Kit

[1] The Ion PGM™ Dx System with Torrent Suite™ Assay Development Software is For Research

Use Only. Not for use in diagnostic procedures.

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Intended use

The Ion PGM™ Sequencer performs real‑time measurements of hydrogen ions

produced during DNA replication.

Library kit compatibility

This sequencing kit can be used with all library types prepared using any Ion library

kit.

Template kit compatibility

The Ion PGM™ Hi‑Q™ View Sequencing Kit is compatible with the following template

preparation kit:

• Ion PGM™ Hi‑Q™ View OT2 Kit (Cat. No. A29900)

Select this template kit when creating a Planned Run.

Software and instrument system compatibility

The procedures in this guide are compatible with the following software and

instrument systems:

• Torrent Suite™ Software version 5.0.4 or later on the Ion PGM™ System. To update

the software, see the Ion PGM™ System Reference Guide. (Pub. No. MAN0009783),

available at thermosher.com.

•Torrent Suite™ Assay Development Software version 5.0.6 or later for Research

Use Only experiments on the Ion PGM™ Dx System[2]. To update the software, see

the Torrent Suite™ Assay Development Software Help. To use this kit on the Ion

PGM™ Dx System with Torrent Suite™ Assay Development Software, see the

following user guides:

–Ion PGM™ Hi‑Q™ View OT2 Kit—Assay Development Mode User Guide (Pub.

No. MAN0015753) and

–Ion PGM™ Hi‑Q™ View Sequencing Kit—Assay Development Mode User Guide

(Pub. No. MAN0015792)

[2] The Ion PGM™ Dx System with Torrent Suite™ Assay Development Software is For Research

Use Only. Not for use in diagnostic procedures.

Chapter 1 Product Information

Intended use

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Kit contents and storage

The Ion PGM™ Hi‑Q™ View Sequencing Kit (Cat. No. A30044) includes the following

components:

Component Amount Storage

Ion PGM™ Sequencing Supplies (Part No. A25587)

Wash 1 Bottle w/ label (250 mL) 1 bottle

15°C to 30°C

Wash 3 Bottle w/ label (250 mL) 1 bottle

Ion PGM™ Reagent Bottle Sipper Tubes (blue) 16 sipper tubes

Ion PGM™ Wash Bottle Sipper Tubes (gray, for

250‑mL bottles)

8 sipper tubes

Ion PGM™ Wash Bottle Sipper Tubes (gray, for 2-L

bottles)

4 sipper tubes

Reagent Bottles w/ labels (50 mL) 25 bottles

Ion PGM™ Hi‑Q™ View Sequencing Reagents (Part No. A30043)

Ion PGM™ Hi‑Q™ View Sequencing Polymerase

(yellow cap)

36 µL

–30°C to –10°CIon PGM™ Hi‑Q™ View Sequencing Primer (white cap) 144 µL

Ion PGM™ Hi‑Q™ View Control Ion Sphere™ Particles

(clear cap)

60 µL

Ion PGM™ Hi‑Q™ View Sequencing Solutions (Part No. A30275)

Ion PGM™ Hi‑Q™ View Sequencing W2 Solution 4 × 125 mL

15°C to 30°C[1]

Ion Cleaning Tablet 4 tablets

Annealing Buffer 12 mL

Ion PGM™ Hi‑Q™ View Sequencing W3 Solution 2 × 100 mL

Ion PGM™ Hi‑Q™ Sequencing dNTPs (Part No. A25590)

Ion PGM™ Hi‑Q™ Sequencing dGTP (black cap) 80 µL

–30°C to –10°C

Ion PGM™ Hi‑Q™ Sequencing dCTP (blue cap) 80 µL

Ion PGM™ Hi‑Q™ Sequencing dATP (green cap) 80 µL

Ion PGM™ Hi‑Q™ Sequencing dTTP (red cap) 80 µL

[1] Store Ion PGM™ Hi‑Q™ View Sequencing W2 Solution protected from light.

Chapter 1 Product Information

Kit contents and storage

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Ion PGM™ Wash 2 Bottle Kit

The Ion PGM™ Wash 2 Bole Kit (Cat. No. A25591) is sold separately, and includes the

following components:

Component Amount Storage

Wash 2 Bottle w/ label (2 L)

Note: Must be conditioned at least 8 hours before

use as described in “Condition the Wash 2 Bottle

for ﬁrst use“ on page 21

1 bottle

15°C to 30°C

Wash 2 Bottle Conditioning Solution 1 × 125 mL

Ion Chip kits

The following Ion Chip kits are compatible with this sequencing kit, and are sold

separately:

Component Amount Catalog No. Storage

Ion 318™ Chip v2 BC 4 pack

8 pack

4488146

4488150

15°C to

30°C

Ion 316™ Chip v2 BC 4 pack

8 pack

4488145

4488149

Ion 314™ Chip v2 BC 8 pack 4488144

Instruments and server

Components Cat. No.

Ion PGM™ System and accessories 4462921

Ion PGM™ Torrent Server 4483643

Ion Chip™ Minifuge:

120 VAC

230 VAC

4479672

4479673

Chapter 1 Product Information

Ion PGM

™

Wash 2 Bottle Kit

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Required materials not supplied

Unless otherwise indicated, all materials are available through thermosher.com.

MLS: Fisher Scientic (sherscientic.com) or other major laboratory supplier.

Note: The procedures in this guide have been veried using these specic materials.

Substitution may adversely aect system performance.

3Item Source

Tank of compressed nitrogen (grade 4.5, 99.995% or better) (see

“Gas cylinders“ on page 15)

MLS

Multistage (dual-stage) gas regulator (0-50 PSI, 2-3 Bar output) Fisher Scientiﬁc

NC0393866 or MLS

Choose from one of the following systems:

ELGA™ PURELAB™ Flex 3 Water Puriﬁcation System

Equivalent 18-MΩ water puriﬁcation system

4474524

MLS

Microcentrifuge (capable of >15,500 × g, ﬁts 1.5‑mL and 0.2-mL

microcentrifuge tubes)

MLS

0.22‑µm or 0.45‑µm vacuum ﬁltration system and ﬁlters (nylon or

PVDF ﬁlters, 1‑L volume)

MLS

Rainin™ Pipet-Lite™ LTS Pipette L‑100XLS 10–100 μL[1]

(Alternatives from Gilson and Eppendorf may be used)

Rainin 17014384

Rainin™ Pipet-Lite™ LTS Pipette L‑20XLS 2–20 μL[1]

(Alternatives from Gilson and Eppendorf may be used)

Rainin 17014392

Rainin™ LTS pipette tips, 200 µL, SR-L200F

(Alternatives from Gilson and Eppendorf may be used)

Rainin 17005859

Rainin™ LTS pipette tips, 20 µL, SR-L10F[1]

(Alternatives from Gilson and Eppendorf may be used)

Rainin 17005860

PCR tubes, Flat Cap, 0.2-mL (do not use polystyrene tubes) Fisher Scientiﬁc

14-222-262

Vortexer with a rubber platform MLS

Thermal cycler with a heated lid MLS

Graduated cylinders (1 L or 2 L volume) MLS

Glass bottle (1 L) MLS

15-mL conical tubes MLS

NaOH (10 M), molecular biology grade MLS

Pipette set and ﬁltered tips, P2, P20, P200, and P1000 μL MLS

Microcentrifuge tubes, 1.5-mL or 1.7-mL MLS

Chapter 1 Product Information

Required materials not supplied

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

3Item Source

Syringe, 10 CC, Female Luer-Lok™ (used for clearing lines) Provided with the

Ion PGM™

Sequencer, or MLS

Ion PGM™ 2.5 L Waste Bottle[2] 4482565

Optional materials

Ion PGM™ Controls Kit v3 [3] A30046

Ion PGM™ Sequencing Sippers Kit[4] 4478682

Non-interruptible Power Supply (UPS)[5] MLS

[1] Ensure tips from any vendors are low binding tips. Required for loading samples onto the Ion PGM™ Chips.

[2] Required one-time purchase for older ("Orange") versions of the Ion PGM™ System.

[3] Not commonly needed, but available for troubleshooting.

[4] Contains additional sipper tubes; not commonly needed.

[5] We recommend using an non-interruptible power supply (UPS) for laboratories that experience frequent

power outages or line voltage fluctuations. The UPS must be compatible with 1500 W output or higher. The

1500 VA unit from APC provides ~11 minutes of backup power for an Ion PGM™ System.

Chapter 1 Product Information

Required materials not supplied

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Ion PGM™ System layout

789

1Touchscreen

2Chip clamp

3Grounding plate

4Power button

5Reagent Bottle Sipper Tube

6Ion Chip™ Minifuge

7Barcode Scanner

8Collection tray

9Reagent Bottle

10 Waste Bottle

11 Wash 2 Bottle (W2 position)

12 Wash 3 Bottle (W3 position)

13 Wash 1 Bottle (W1 position)

Chapter 1 Product Information

Ion PGM

™

System layout

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Before you begin

For additional safety information, see Appendix D, “Safety“.

IMPORTANT! Before proceeding, make sure that you have updated the Torrent

Suite™ and Ion PGM™ System software to version 5.0.4 or later. See the Ion PGM™

System Reference Guide (Pub. No. MAN0009783) for more information.

IMPORTANT! The Ion PGM™ System is installed by trained service personnel and

must not be relocated without assistance from trained service personnel. See

“Customer and technical support“ on page 64.

IMPORTANT! A primary source of contamination is DNA fragments from previously

processed samples. Do not introduce amplied DNA into the library preparation

laboratory or work area.

IMPORTANT! Possible contamination can occur during the transfer of dNTPs into

Reagent Tubes. Be careful to avoid cross contamination of dNTP stocks. Barrier tips

are required for all pipeing steps. Change gloves after handling concentrated dNTP

stocks.

IMPORTANT! Dry ice (solid CO2) must be kept away from areas where buers, wash

solutions, or sources of molecular biology grade water for the Ion PGM™ System are

used. High air concentrations of subliming CO2 may change the pH of such buers

during or after their preparation. The stability of the pH of these buers is a critical

factor in the performance of the Ion PGM™ System.

IMPORTANT! Signicant vibration during sequencing may add noise and reduce the

quality of the measurements. The Ion PGM™ System must be installed on a bench that

is free from vibrations or in contact with equipment that can cause vibrations to the

bench (freezers, pumps, and other similar equipment).

IMPORTANT! Position the Ion PGM™ System so that the front bezel is a minimum of

12 in. (30.5 cm) and the Reagent Tubes containing dNTPs are a minimum of 8 in.

(20.3 cm) from the front of the laboratory bench. Place the instrument at least 40 in.

(1 meter) away from major sources of electronic noise such as refrigerators or

microwaves.

Update the

software

Instrument

installation by

trained personnel

only

Nucleic acid

contamination

CO2 contamination

Instrument

vibration and

clearances

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

IMPORTANT! To avoid possible damage to the chip from static electricity, prior to

handling chips, you must ground yourself on the grounding plate (located next to the

chip clamp) by touching the grounding plate with your bare hand.

Do not place chips on non‑grounded surfaces such as a bench.

Always use the grounding plate to hold chips that are not in the package inserted in

the chip clamp or the Ion Chip™ Minifuge bucket.

WARNING! Instrumentation must be installed and operated in a well‑

ventilated environment, dened as having a minimum airow of 6–10 air

changes per hour. Assess the need for ventilation or atmospheric monitoring to

avoid asphyxiation accidents from inert gases and/or oxygen depletion, and

take measures to clearly identify potentially hazardous areas through training

or signage. Please contact your Environmental Health and Safety Coordinator

to conrm that the instruments will be installed and operated in an

environment with sucient ventilation.

Guidelines for sequencing runs

•When performing two sequencing runs from the same initialization, the second

run must be started within 24 hours after start of initialization.

• If you press the Abort buon on the sequencer touchscreen, the touchscreen may

freeze. You may need to restart the sequencer.

IMPORTANT! After aborting a run, do not open the chip clamp, reagent tubes, or

wash boles until a new run or cleaning is initiated. Doing so can cause a uid or gas

leak if the sequencer was in a pressurized state when the run was aborted. From the

main menu, select either Clean or Run, then follow the touchscreen prompts to

depressurize the system.

Guidelines to prevent cross-contamination

•Use good laboratory practices to minimize cross‑contamination of products and

reagents.

• When designing the laboratory layout, consider separating pre‑ and post-

amplication activities. To signicantly reduce the potential for contamination,

dedicate laboratory supplies and/or equipment to the appropriate space.

Static electricity

Ventilation

requirements

Chapter 2 Before you begin

Guidelines for sequencing runs

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Guidelines for pipetting

• Vortex all reagents except enzymes for ~5 seconds. Mix enzymes by icking the

tube with your nger 4 times. Briey centrifuge to collect the contents before use.

• Ensure that all reagents are completely thawed at room temperature, such that no

ice crystals are visible in the tube.

• Pipet viscous solutions slowly and ensure complete mixing.

• Change tips between pipeing steps.

Guidelines for chip handling and use

• Always remove gloves before transferring chips onto or o the instrument. Hold

chips by their edges when handling.

• To avoid damage due to electrostatic discharge (ESD), do not place chips directly

on the bench or any other surface. Always place chips either on the grounding

plate on the sequencer or in the Ion Chip™ Minifuge bucket.

• Used chips cannot be reused for sequencing. Used chips must be discarded or

clearly marked for cleaning and initialization.

Gas cylinders

You must supply the required nitrogen gas cylinder and accessories for the

installation. This instrument requires a pressurized house line or one size 1‑A

nitrogen gas cylinder that holds approximately 7.2 m3 (257 ft3) of gas when full. Use

only prepuried nitrogen of 99.995% (grade 4.5) or greater purity.

CAUTION! Damage to the instrument and its products can result from using

impure gas, gases other than nitrogen, or an inadequate amount of gas.

WARNING! EXPLOSION HAZARD. Pressurized gas cylinders are potentially

explosive. Always cap the gas cylinder when it is not in use, and aach it rmly

to the wall or gas cylinder cart with approved brackets or chains.

WARNING! Gas cylinders are heavy and may topple over, potentially causing

personal injury and tank damage. Cylinders should be rmly secured to a wall

or work surface. Please contact your Environmental Health and Safety

Coordinator for guidance on the proper installation of a gas cylinder.

Chapter 2 Before you begin

Guidelines for pipetting

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

To perform a leak test on the gas cylinder:

1. Open the main tank shuto valve. The high‑pressure gauge of the gas tank

regulator reads approximately 2,000–2,500 psi for a full tank.

2. Adjust the pressure to the instrument by slowly turning the pressure adjustment

valve clockwise until the low‑pressure gauge reads 30 psi.

3. Close the needle valve, then close the main tank valve.

4. Monitor the high‑pressure gauge of the gas tank regulator for 5 minutes. There

should be no noticeable drop in pressure.

If the pressure Action

Drops in 5 minutes There can be a leak at either the needle valve or the gas tank

regulator itself. Check the ﬁttings and resolve any problems,

then continue with step 5.

Does not drop in

5 minutes

The instrument passes the leak test. Reopen the main tank

valve and skip the following steps.

5. Open the main tank valve and the needle valve for at least 15 seconds to

pressurize the instrument.

6. Close the main tank valve.

7. Monitor the high‑pressure gas tank regulator gauge. There should be no more

than a 100‑psi drop in pressure after 5 minutes. Locate, then resolve any leaks.

Turn the main tank valve back on.

Perform a leak

test

Chapter 2 Before you begin

Gas cylinders

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Protocol workﬂow

Clean the sequencer

Yes No

Create a

Planned Run

Initialize the

Sequencer

Load the chip

and sequence

Reinitialization

required before

the next run?

Chapter 2 Before you begin

Protocol workﬂow

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Create a Planned Run

Planned Runs can be created with the following software programs for use on the

following systems:

Software Instrument System

Torrent Suite™ Software, v5.0.4 or later Ion PGM™ System

Torrent Suite™ Assay Development Software, v5.0.6

or later

Ion PGM™ Dx System

IMPORTANT! If you are using the Ion PGM™ Dx System with Torrent Suite™ Assay

Development Software[3], see the following user guides:

·Ion PGM™ Hi‑Q™ View OT2 Kit—Assay Development Mode User Guide (Pub. No.

MAN0015753) and

·Ion PGM™ Hi‑Q™ View Sequencing Kit—Assay Development Mode User Guide (Pub.

No. MAN0015792)

Planned Run set‑up and the Ion PGM™ Dx System workow dier from those

described in this user guide.

About Planned Runs

Planned Runs contain all the seings used in a sequencing run, including number of

ows, kit types, barcodes, sample information, and reference les (if any). They are

used to track samples, chips, and reagents throughout the sequencing workow, from

template preparation through sequencing and subsequent data analysis.

You can create a Planned Run in the Torrent Suite™ Software or Torrent Suite™ Assay

Development Software on the Torrent Server connected to your sequencer, and then

select the appropriate plan on the sequencer touchscreen when you start the run.

You can also create a Planned Run on one Torrent Server and then transfer it to

another server for sequencing. See “Enable Planned Run sharing“ on page 55 for

more information.

[3] The Ion PGM™ Dx System with Torrent Suite™ Assay Development Software is For Research

Use Only. Not for use in diagnostic procedures.

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

Create a Planned Run

The following provides a summary of steps for creating a Planned Run in Torrent

Suite™ Software, for use on the Ion PGM™ System.

For more detailed instructions, see the Torrent Suite™ Software Help Guide, available at

thermosher.com/ion-pgm-docs.

1. Open the Torrent Browser on a computer connected to your sequencing system.

2. Select the Plan tab, then select Templates.

3. Select the application in the left navigation bar (for example, AmpliSeq DNA). A

list of existing Planned Run templates for that application will be displayed.

Select one of the following options to create a new plan:

• To create a new Planned Run without using an existing template, click on

Plan New Run.

• To create a new Planned Run from an existing template, click the buon

for the template and select Plan Run from the drop‑down menu.

• Other options may be available depending on the selected application, such

as downloading templates from AmpliSeq.com.

Chapter 3 Create a Planned Run

Create a Planned Run

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

4. In the wizard, make your selections on each screen, then click Next to proceed to

the next screen.

Note: For a complete description of each option, see the Torrent Suite™ Software

Help Guide.

5. When you have completed your selections, click Plan Run.

The run will be listed on the Planned Runs screen under the name you specied,

and is available on the sequencer when you are seing up the run.

Chapter 3 Create a Planned Run

Create a Planned Run

Ion PGM

™

Hi‑Q

™

View Sequencing Kit User Guide

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