Ion ReproSeq™ PGS 316 Kit, without chips - FAQs

The maximum number of Ion SingleSeq libraries that can be pooled and sequenced on a chip depends on the chip being used and can vary between 2-24 samples/run:

- Ion 314 Chip Kit v2 (BC or non-BC) = 2 libraries per chip
- Ion 316 Chip Kit v2 (BC or non-BC) = 16 libraries per chip
- Ion 318 Chip Kit v2 (BC or non-BC) = 24 libraries per chip

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The enriched ISPs can be stored at 2-8 degrees C for up to 3 days.

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The template ISPs should be in the Ion PGM Template IA Wash Solution and can be stored at 4 degrees C for up to one week.

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Diluted library pools and non-diluted library pool stock solutions may be stored for 1 week at 4 degrees C. After 1 week, we recommend that you thaw the individual Ion SingleSeq libraries and repeat the pooling, purification, quantification, and dilution steps starting from the “Pool the libraries“ step in the protocol.

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Sample libraries may be stored individually at -30 degrees C to -10 degrees C before proceeding to the next step. We do not recommend storing library pools prior to quantification.

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Purified gDNA can be used with the Ion ReproSeq PGS Kit and the recommended amount is 15-60 pg.

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1-10 cells are recommended for use with the Ion ReproSeq PGS Kit.

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Do not exceed 4 freeze/thaw cycles.

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If only a subset of barcodes is needed, Thermo Scientific the foil of the barcode adapter to collect the required amount of barcode adapter for the reaction. If you are using a subset, after piercing, seal the Thermo Scientificd wells with laboratory tape to minimize cross-contamination.

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Yes, the Ion SingleSeq Kit (Cat. No. A28955, 24 reactions/kit), or Ion SingleSeq 96 Kit (Cat. No. A34763, 96 reactions/kit) are available separately.

The Ion ReproSeq PGS Kits - Ion S5 System, are also available separately and include reagents to prepare templates and load 4 chips on the Ion Chef Instrument, and sequence the chips on the Ion S5 or Ion S5 XL Sequencer.
- Ion ReproSeq PGS Kit with Ion 520 Chips (24 samples/run), Cat. No. A34900
- Ion ReproSeq PGS Kit with Ion 530 Chips (96 samples/run), Cat. No. A34901

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Each barcode is intended for single use.

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The Ion ReproSeq PGS Kit workflow does not use the Ion OneTouch 2 System for template preparation. Template preparation is performed using Ion IA technology, whereby DNA is clonally amplified onto a bead surface through a non-emulsion, isothermal reaction.

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Six configurations of the Ion ReproSeq PGS Kit are available. The kits vary in the number of Ion SingleSeq kits provided, the maximum number of samples processed per kit, the type of Ion sequencing chip provided, and whether Ion sequencing chips are included. Depending upon the configuration, there are 4 or 5 subkits in the Ion ReproSeq PGS kits:

- Ion SingleSeq Kit includes reagents to extract, amplify, and barcode genomic DNA.

- Ion PGM Template IA 500 Kit includes reagents for preparing template-positive Ion PGM Template IA Ion Sphere Particles (ISPs) for sequencing with the Ion PGM System.

- Ion PGM Hi-Q Sequencing Kit includes reagents and materials for the initialization and sequencing runs on the Ion PGM Sequencer.

- Ion PGM Hi-Q Wash 2 Bottle Kit

- Ion Chip Kit v2: The Ion ReproSeq PGS kits are available with three different types of Ion Chips Kits v2: Ion 314 Chip Kit v2, Ion 316 Chip Kit v2, and Ion 318 Chip Kit v2. The Ion Chips Kits v2 in the Ion ReproSeq PGS kits are barcoded (BC). The non-barcoded (non-BC) versions of the Ion Chip Kits v2 are also available and can only be purchased as standalone items.

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The Ion ReproSeq PGS workflow takes approximately 8-10 hours depending on the Ion Chip type in use. The library construction takes ˜3.5 hours and the template preparation takes 2 hours. The run time for an Ion 314 Chip takes 2.5 hours, an Ion 316 Chip takes 3.5 hours, and an Ion 318 Chip takes 4.5 hours.

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The Ion ReproSeq PGS (Preimplantation Genetic Screening) kits are bundles of four or five sub-kits which, when used in conjunction with the Ion PGM System, provide reagents and materials for whole genome amplification and sequencing to detect chromosomal aneuploidies, chromosome arm events (>48 Mb), and copy number variations starting from a single cell.

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