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查看更多产品信息 Ion PGM™ Hi-Q™ View OT2 Kit - FAQs (A29900)
30 个常见问题解答
The "time remaining" is not entirely accurate. The centrifuge may keep rotating for about 15 minutes after the time has become 0:00. If it rotates for more than 15 minutes, please contact Technical Support.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
The pressure inside the instrument may be above normal. If the Ion OneTouch 2 firmware is older than v540, please update it to the latest version.
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It may be rubber from the packing. The amount varies among instruments and it is not anything to be concerned about, as it will be removed by the Ion OneTouch ES.
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Firmware updates to the software controlling the Ion OneTouch 2 Instrument are periodically released. To update the firmware to the appropriate version, use either a USB flash drive or an ethernet connection, as described on page 11 of the Ion OneTouch 2 System - Installation, Setup, and Maintenance User Guide (http://tools.thermofisher.com/content/sfs/manuals/MAN0014388_IonOneTouch2Sys_UG.pdf ).
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
For the Ion PGM System, we offer:
- Ion PGM Hi-Q View OT2 Kit (Cat. No. A29900) for template preparation of libraries up to 400 bp in length.
For the Ion Proton System, we offer:
- Ion PI Hi-Q OT2 200 Kit (Cat. No. A26434) for template preparation of libraries up to 200 bp in length.
For the Ion S5 System and Ion S5 XL System, we offer two kits:
- Ion 520/530 Kit-OT2 (Cat. No. A27751) for template preparation of libraries up to 400 bp in length.
- Ion 540 Kit- OT2 (Cat. No. A27753) for template preparation of libraries up to 200 bp in length.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
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.
We recommend using T Cell Leukemia (Jurkat) Total RNA (Cat. No. AM7858) as a control. T cell Leukemia (Jurkat) Total RNA is derived from a cell line consisting of a single T cell clonotype. Running the Oncomine TCR Beta-LR Assay on Jurkat Total RNA should enable detection of a single clonotype.
Unlike other AmpliSeq panels, there are no BED files and no reference is needed. All downstream analysis takes place in Ion Reporter Software.
We have tested the Ion Library Equalizer Kit with this kit for library quantitation and it does function, but manually balancing library input into the Ion Chef instrument is preferred.
Torrent Suite Software 5.6 and Ion Reporter Software 5.6 are needed to generate reports. In Torrent Suite Software 5.6, two Planned Run templates are provided for use with the Oncomine TCR Beta-LR Assay.
The Oncomine TCR Beta-LR Assay has been developed and verified for use on the Ion GeneStudio S5 Series System. The assay can be used with the Ion PGM System, however, this instrument configuration is not supported.
Here are the components of the Oncomine TCR Beta-LR Assay:
- Oncomine TCR Beta‑LR Panel
- Ion AmpliSeq Library Kit Plus
- Ion Select Barcode Adapters
The primer pool is at 5X concentration.
The Oncomine TCR Beta-LR Assay enables long-read RNA sequencing (up to 400 bp amplicon length) for complete characterization of the T cell receptor Beta (TCR Beta) chain, including all three complementarity-determining regions (CDR1, 2, and 3) of the variable gene. This enables allele-specific interrogation of TCR Beta to potentially predict drug-mediated adverse events.
The primers are provided as a single pool.
Up to 16 samples can be multiplexed on an Ion 530 chip, up to 4 samples can be multiplexed on an Ion 520 chip, and up to 4 samples can be multiplexed on an Ion 318 chip.
A variety of research sample types including fresh-frozen tissue, whole blood, and sorted T cells are compatible with the Oncomine TCR Beta-LR Assay.
T cell repertoire sequencing helps:
- To characterize diversity and study features of T lymphocytes in blood and infiltrated tumors
- To study variable gene polymorphisms implicated in autoimmune disease or immune-mediated adverse events
- To study the production and function of therapeutic or engineered T cells
This kit is good for 24 samples.
The Oncomine TCR Beta-LR Assay is an RNA-based next-generation sequencing (NGS) research assay that enables the characterization of the T cell receptor Beta (TCR Beta) sequences including all three complementarity-determining regions (CDR1, 2, and 3) of the variable gene. The assay measures T cell repertoire diversity as well as clonal expansion, and allows for identification of allele-specific polymorphisms.
No, the Ion PGM Hi-Q View OneTouch 2 Kit has only been validated with the Ion PGM Hi-Q View Sequencing Kit.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
In addition to updating the Ion OneTouch 2 System firmware to Torrent Suite Software v.5.0.4 or later, you will have to perform the chemistry change protocol. To set up the Ion OneTouch 2 Instrument when switching between chemistries, refer to Chapter 5 of the Ion PGM OneTouch 2 Hi-Q View user guide,“Demonstrated Protocol: Setting up the Ion OneTouch 2
Instrument when switching between template preparation kits”.
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You will need Torrent Suite Software v5.0.4 or later on the Ion PGM System.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
Yes, the Ion PGM Hi-Q View chemistry is compatible with both 200 bp and 400 bp libraries.
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Yes, the Ion PGM Hi-Q View sequencing chemistry is compatible with Ion ReproSeq PGS kits.
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Visible Ion Sphere particle (ISP) beads help minimize user variability in manual chip loading, to help improve experimental reproducibility and confidence in your results. Additionally, the new Ion Sphere particles have been optimized to provide increased sequencing performance.
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The calibration standard is an optional reagent and is recommended for samples that have no reference or small references. It is a small panel consisting of known sequence containing homopolymers. It provides a reference for base calibration but the option needs to be selected in run planning.
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The Ion PGM Hi-Q View OneTouch 2 Kit includes the reagents required for preparing 8 reactions of template-positive Ion PGM Hi-Q View Ion Sphere Particles (ISPs) on the Ion OneTouch 2 System. The components are:
- Ion PGM View OneTouch 2 Supplies
- Ion PGM Hi?Q View OneTouch 2 Reagents
- Ion PGM Hi?Q View OneTouch 2 Solutions
- Ion PGM Calibration Standard
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
In a typical Ion OneTouch 2 run, you should expect ~18 mm drop in the level of oil in the tube (~22 mL consumed), and ~13 mm drop in the recovery solution (~15 mL consumed). During cleaning, you should expect to collect ~10 mL of oil in the 50 mL conical waste collection tube.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
Ion Torrent OneTouch2 protocols can be found at our NGS Support Center (thermofisher.com/ngssupport), in the NGS Template Preparation Support Center section. For all other protocols, please go here (http://www.thermofisher.com/manuals-protocols-search.html).
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.