Ion Personal Genome Machine™ (PGM™) System - FAQs

View additional product information for Ion Personal Genome Machine™ (PGM™) System - FAQs (4462921)

36 product FAQs found

What should I do if W1 fails at Raw Traces and the screen turns red during the last step of initialization of my Ion PGM System?

The pH of W1 may be out of range or the volume of W1 may be too small or there may have been a minor problem during measurement. Check the amount of W1, press the 'Start' button on the screen, and restart the measurement. If it passes, continue to start the run. If an error appears again, please note the pH of W1, the amount of W1, as well as the error message, and contact Technical Support.br/>
Please note that passing pH values for the reagents may vary depending on the sequencing kit used, but the instrument will automatically determine the correct pH range depending on the sequencing kit selected during initialization. If you are performing the reagent check outside of the initialization, please contact Technical Support for guidance.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Are libraries generated with the original Ion Total RNA-Seq Kit (Cat. No. 4466666) compatible with the Ion PGM and Proton systems?

Yes, RNA-Seq libraries prepared using the Ion Total RNA-Seq Kit are compatible with the current Ion PGM and Ion Proton systems.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

What is the thermal output of the Ion PGM System?

The Ion PGM System typically draws ~200 to 300 W and outputs ~682 to 1023 BTU/h.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

At what pressure should I set the gas regulator for my Ion PGM Sequencer? When should I change out my tank?

Please ensure that the gas pressure going to your instrument is set to 30 PSI. Monitor the pressure in your tank closely, and exchange with a fresh tank when the pressure goes down to 500 PSI.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How do I update my Ion PGM to the latest software version?

Firmware updates for Ion instruments are included with the latest version of Torrent Suite Software. First, download and install the latest version of Torrent Suite Software on your Torrent Server. Once completed, your Ion PGM System should alert you that an update is available. You can also start the update process from the Options menu. From the Main menu, select Options, then Updates, Released Updates, and Check. Once your PGM system has detected the update, press the Update button to begin the process. Instructions to update the Ion PGM Sequencer can also be found in the Appendix of the Ion PGM Sequencing kit user guides.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How do I properly shut down my Ion PGM System?

It is not necessary to power off the instrument overnight or over the weekend. If the instrument will not be used for more than 3 days, the instrument may be powered off.

To power off the instrument:
1) In the Main Menu, select Tools > Shut Down.
2) If you have not already cleaned the instrument, select 18 MΩ water cleaning, then press Next to start the cleaning process.
3) When cleaning is complete, press Shut Down.
4) After you exit the main touchscreen, press the Halt button, then OK when prompted. The instrument will power down.

Instructions to power the Ion PGM Sequencer on or off can also be found in the Appendix of the Ion PGM Sequencing kit user guides. We recommend performing a chlorite cleaning prior to the instrument's next use.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How long does a nitrogen tank typically last?

A size 1–A tank typically lasts for ~4–6 weeks with consistent usage.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

May I use consumables not listed in the Ion PGM Sequencing kit user guides?

Protocols have been validated using the consumables listed in the user guides. For optimal performance and complete support, we recommend using all items as listed.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How should I store my Ion PGM Sequencing kit reagents?

Proper storage conditions are provided in the sequencing user guide under the “Kit Contents and Storage” section.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Where can I find the Ion PGM System protocols for my kit?

Ion PGM System user guides and quick references can be accessed and downloaded from the Next Generation Sequencing Support Center (thermofisher.com/ngssupport), in the Library Preparation, Template Preparation, Sequencing, or Software sections.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

What read lengths are supported by the Ion PGM System?

Currently, the Ion PGM System supports up to 400–base reads.

For up to 400-base reads, using the Ion PGM Hi-Q View Chemistry and Ion PGM One Touch 2 system, we recommend:
• Ion PGM Hi-Q View One Touch 2 Kit (Cat. No. A29900)
• Ion PGM Hi-Q View Sequencing Kit (Cat. No. A30044)

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How does the Ion Torrent technology work?

Ion Torrent semiconductor sequencing is the first post-light sequencing technology. Ion sequencing works by taking advantage of the natural release of hydrogen ions during nucleotide incorporation by DNA polymerase. By directly coupling chemical and digital information, Ion Torrent platforms provide a fast, simple, and scalable sequencing solution to meet your lab's needs.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Can I update Torrent Suite server without internet access?

Yes, you can update it, but a service engineer would have to be called onsite to carry it out.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

I have reanalyzed a sequencing run in Torrent Suite Software and it has completed with 96 errors, what can I do?

You should reanalyze from base calling. If this fails, please contact your local Bioinformatics support.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How do I change a user's password in Torrent Suite Software?

Refer to page 313 of the Torrent Suite Software 5.8 manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017530_TorrentSuite5.8_UG.pdf) to change a user's password.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Which format should I use to create a hotspot file for the variant caller in Torrent Suite Software?

Formats allowed are explained on page 258 of the Torrent Suite Software 5.8 manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017530_TorrentSuite5.8_UG.pdf).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Can I install Torrent Suite Software outside the Torrent server?

Yes, you can get the software from https://github.com/iontorrent/TS and install in another server. Additionally, you can download the TMAP aligner and Torrent variant caller used in TSS, among other software.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Where can I find latest changes for Torrent Suite Software's release?

You can find the main changes in each version's release notes available from www.thermofisher.com. For example, release notes from Torrent Suite Software 5.8 version can be found at https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017478_TorrentSuiteSoftware_5_8_ReleaseNotes.pdf. Alternatively, you can look for “torrent suite software release notes” using your search engine.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How can I customize variant caller parameters in Torrent Suite Software?

A description of parameters in Torrent browser is provided on page 225 of the Torrent Suite Software 5.8 manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017530_TorrentSuite5.8_UG.pdf).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Why was my variant not called in Torrent Suite Software?

Filtered variants can be found as explained on page 206 of the Torrent Suite Software 5.8 manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017530_TorrentSuite5.8_UG.pdf).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Can I downgrade Torrent Suite Software on my own?

No, it is not possible to downgrade Torrent Suite Software after upgrade.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Where I can find unaligned reads (uBAM) on Ion S5 System's run report?

Unaligned reads are merged with aligned reads within the output BAM files whenever a reference genome is provided. The unaligned BAM files are deleted to save space.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

What are the types of next-generation sequencing data files and which should be archived?

Signal Processing Input - Required to reanalyze an Ion Torrent Ion PGM or Ion Torrent Ion Proton run from scratch. (The Output file category is also required).

Basecalling Input - Required to re-analyze an Ion PGM or Ion Proton run from basecalling. (The Output file category is also required).

Output - Required to see the run report in the Torrent Browser and either to reanalyze the run or to launch a plugin.

Intermediate - Useful to troubleshoot a run. Not required for the run report or for re-analysis.

Intermediate files can be deleted if not needed for run troubleshooting. Depending on your institution, you may need to keep all other files (Signal Processing, Basecalling, and Output).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How do I change the barcoding set used to analyze a next-generation sequencing run?

Follow these steps to change the DNA barcode ID:

- Open a run reports listing in the “Data,” “Completed Runs & Reports” tab.
- In the list view, click the “Edit” button on the right side of the run listing. The Edit Run window opens.
- Click the barcode menu pull-down and choose the correct barcode set. If you have custom barcode sets, your list is different. Click the “Save” button (in the bottom right corner).
- You can now reanalyze the run from the “Data,” “Completed Runs & Reports” tab.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How do I reanalyze my next-generation sequencing data with a different reference sequence?

Log in to Torrent Browser and click the “Data” tab, “Completed Runs & Reports page”, and find your run name. In the table view, click the “Reanalyze” option in the gear menu on the right of the run entry. The main run analysis dialog opens. Click the “Advanced” button to display the additional run parameter options. Click the “Alignment Reference” pull-down menu and select the reference for this run. (Optional) Change other advanced options if required. (See “Work with Completed Runs” for a description of the advanced fields.) Click the “Start Analysis” button.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Why do I see the sequence GGCCAAGGCG at the beginning of the majority of the reads in my next-generation sequencing data analysis?

The GGCCAAGGCG sequence is part of the RNA library adapter sequence. This sequence can be automatically trimmed by the software by selecting the “RNA_Barcode_None” option under the Barcodes menu. The run can be reanalyzed with the correct barcode settings to trim the adapter sequence. In the future, the “RNA_Barcode_None” option should be selected in the run plan or selected on the instrument prior to starting the sequencing run.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How do I archive my Ion Torrent sequencing data?

Please refer to the Data Management section of the Torrent Suite Software user guide for your version of software. The user guides can be found here under Documents: https://www.thermofisher.com/us/en/home/technical-resources/technical-reference-library/next-generation-sequencing-support-center/ngs-software-support.html

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How do I analyze my Ion Torrent sequencing data?

Primary analysis and secondary analysis of the data is performed by the Ion Torrent Server. Depending on the application, there may be tertiary analysis plugins that can be installed on the Torrent Server. For other applications, such as RNA-Seq, we recommend using third-party software. For more information, please see the Ion NGS Analysis Workflow page (https://www.thermofisher.com/us/en/home/life-science/sequencing/next-generation-sequencing/ion-torrent-next-generation-sequencing-workflow/ion-torrent-next-generation-sequencing-data-analysis-workflow.html).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Where can I find the user guides and release notes for Torrent Suite Software?

You can find user guides, release notes, and additional information on the Torrent Suite Software here (https://www.thermofisher.com/us/en/home/technical-resources/technical-reference-library/next-generation-sequencing-support-center/ngs-software-support.html).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

What is the Ion Torrent informatics workflow?

For more information on the informatics workflow and analysis software, such as Ion Reporter and Torrent Suite, please go to the Ion NGS Informatics workflow page (https://www.thermofisher.com/us/en/home/life-science/sequencing/next-generation-sequencing/ion-torrent-next-generation-sequencing-workflow/ion-torrent-next-generation-sequencing-data-analysis-workflow.html).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

I'm having problems using an old, used chip, for initialization of my Ion Proton System. Is there anything I can do?

Rinse the Proton chip by pipetting 100 µL of isopropanol into the chip, followed by 100 µL of water. This may remove any bubbles or reside from the chip surface that may cause initialization issues.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

What should I do if an error message, "Calibration FAILED" appears during initialization of my Ion PGM System?

Open the chip clamp and check for any leaks. If you see solution leaking, please remove the chip and clear the solution with a Kimwipe tissue. DO NOT wipe or scrub, as the socket could be damaged. Clean the socket by gently touching it with a damp Kimwipe tissue. Make sure the socket is dry and replace the chip. Press the “Calibration” button to recalibrate the chip. If it fails again, change the chip to a different one. Also, if you are using an Ion 316 chip v2 on a v3.4 or older instrument, the instrument will not recognize the chip.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

What should I do if a "No Connect FTP Server" alert appears on the Ion PGM touch screen?

The Ion PGM System and Torrent Server may not be connected. Please shut down the system and server and reboot them. The server may enter a system check after the reboot, which will take 3-4 hours. If you have a monitor and keyboard connected directly to the Torren Server, you can avoid the system check by pressing “c” on the keyboard.

If there is a reason you cannot reboot the system, you can run the Ion PGM System without connection to the server. The runs will be saved inside the system. For 200 bp sequencing, here is the number of runs that may be saved for these chip types:

Ion Torrent Ion 314 chip: 40 runs

Ion Torrent Ion 316 chip: 6 runs

Ion Torrent Ion 318 chip: 5 runs

The data will automatically transfer to the server after connection.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

What should I do if R1, R2, R2 and/or R4 fails at Raw Traces and the screen turns red during the last step of initialization of my Ion PGM System?

The pH of the nucleotides may be out of range or there may have been a minor problem during measurement. Press the “Start” button and restart the measurement. If it passes, continue to start the run. If an error appears again, please note the pH of R1, R2, R3, and R4, as well as the error message, and contact Technical Support.

Please note that passing pH values for the reagents may vary depending on the sequencing kit used, but the instrument will automatically determine the correct pH range depending on the sequencing kit selected during initialization. If you are performing the reagent check outside of the initialization, please contact Technical Support for guidance.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

How do I evaluate run performance of an Ion Torrent Ion PGM run?

Please refer to the “How to assess a PGM sequencing run report” in the Documents section of the NGS Sequencing Support Center (https://www.thermofisher.com/us/en/home/technical-resources/technical-reference-library/next-generation-sequencing-support-center/ngs-support.html).

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

For my application, what kits do I need for sequencing on the Ion Torrent Ion PGM System?

Please see the Ion PGM System Product Selection Guide (http://tools.thermofisher.com/content/sfs/brochures/Consumables-Product-Selection-Guide.pdf) for a list of available products for the Ion PGM System workflow organized by application.

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.