Ion Xpress™ Barcode Adapters 1-96 Kit - FAQs

No, the Ion Xpress Barcode adapters are designed with a 3 bp adapter sequence (GAT) at the 3' end of each barcode to help minimize adapter ligation bias. When pooling barcoded libraries, accurate library quantification is extremely important to help ensure even barcode representation on the sequencing run.

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The Ion Xpress Barcode Adapters kits are available as a set of 96 (Cat. No. 4474517) or in sets of 16 (Cat. Nos.4471250, 4474009, 4474518, 4474519, 4474520 or 4474521). Each barcode kit is sufficient for preparing ? 10 libraries per barcode (10 x 16 libraries or 10 x 96 libraries) for 100 ng input, or 2 libraries per barcode for 1 µg input.

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The number of samples that can be multiplexed in a single sequencing run depends on the capacity of the chip, the size of the library, and the required coverage. A table of approximate capacities based on the size of the library and chip can be found in the Ion Ampliseq Preparation User Guide (https://tools.thermofisher.com/content/sfs/manuals/MAN0006735_AmpliSeq_DNA_RNA_LibPrep_UG.pdf) within the section “Strategies for combining Ion Ampliseq libraries.”

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The TAG Sequencing Barcode Set contains adapters designed for Oncomine workflows using AmpliSeq HD technology and they can be used only with those library kits. This type of chemistry uses a molecular tag in order to track polymerase errors and increase the assay sensitivity. The Ion Xpress Barcode Adapters are not an alternative of the TAG Sequencing Barcode Set and vice versa. The list of the barcodes set compatible with a library kit is specified in the library manual.

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We recommend sequencing up to 5 DNA or RNA samples (4 samples and a non-template control (NTC)) on a single Ion 550 Chip.

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The Oncomine Comprehensive Assay Plus, manual library preparation (Cat. No. A46721) is designed to prepare barcoded sample libraries from DNA and RNA. The kit consists of Oncomine Comprehensive Assay Plus DNA (2-pool) (Part No. A45615), RNA Oncomine Comprehensive Assay v3M (2-pool) (Part No. A33637), and two kits of Ion AmpliSeq Library Kit Plus (Cat. No. 4488990). Sufficient reagents are provided to prepare libraries from 24 samples.

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10 ng of nucleic acid input per pool (20 ng of DNA and 20 ng of RNA) isolated from FFPE samples, including fine needle biopsies, is needed for the Oncomine Comprehensive Assay Plus, manual library preparation.

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The assay is optimized for use with formalin-fixed paraffin embedded (FFPE) samples.

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The Oncomine Comprehensive Assay Plus, manual library preparation (Cat. No. A46721) is a targeted, next-generation sequencing (NGS) assay that enables the detection of single nucleotide variants (SNVs), insertions and deletions (INDELs), copy number variations (CNVs), gene fusions, splice variants, tumor mutational burden (TMB), and microsatellite instability (MSI) across 500+ unique genes. This version of the assay is designed for manual library preparation, not for automated library preparation using the Ion Chef System.

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Here are the products you would need to order:

- Ion Xpress Barcode Adapters Kits
- Agencourt AMPure XP (Beckman Coulter Cat. No. A63880 or A63881)
- Ion Library Equalizer Kit (Cat. No. 4482298)