NGS Reverse Transcription Kit - FAQs

The NGS Reverse Transcription Kit is shipped on dry ice and we recommend storing all components at -30 degrees C to -10 degrees C (non-frost-free).

The NGS Reverse Transcription Kit provides enough material to perform 32 reactions based on 10 µL reactions.

The NGS Reverse Transcription Kit is the first cDNA synthesis kit developed specifically for next-generation sequencing (NGS) applications with specific validated assays. The kit contains 2 components:
- The 10X RT Enzyme Mix includes SuperScript IV Reverse Transcriptase, a proprietary RNase inhibitor, with helper proteins, and stabilizer proteins (1 tube x 32 µL)
- The 5X Reaction Buffer contains dNTPs, random hexamers, and MgCl2 in a formulation optimized for NGS libraries (1 tube x 64 µL)

We recommend using up to 2.5 µg of total RNA as starting material in a 10 µL reaction.
Note:
- Input amounts and reaction volumes vary with each assay and sample type. Consult assay-specific user guides for recommendations.
- To avoid PCR inhibition, RT reaction volume should be equal to or less than 50% of the PCR reaction.

A variety of sample types can be used with the NGS Reverse Transcription Kit, including RNA from blood, bone marrow, FFPE, and partially degraded samples. The NGS Reverse Transcription Kit is compatible with Ion AmpliSeq and Ion AmpliSeq HD assays and can be used with both manual and automated workflows.

For the Ion 530 Chip, 8 libraries can be loaded. For the Ion 540 Chip, the number of libraries that can be loaded ranges from 8 to 32, depending on the size of the libraries. For the Ion 550 Chip, the number of libraries that can be loaded ranges from 12 to 48.

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

The Ion AmpliSeq Mouse TCR Beta SR Assay, RNA is compatible with the Ion 530 Chip, Ion 540 Chip, and Ion 550 Chip.

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

The Ion AmpliSeq Mouse TCR Beta SR Assay, RNA (Cat. No. A45489) consists of the Ion AmpliSeq Mouse TCR Beta-SR Panel, the Ion AmpliSeq Library Kit Plus (Cat. No. 4488990), and the Ion Torrent NGS Reverse Transcription Kit (Cat. No. A45003). Sufficient reagents are provided for the rapid preparation of 24 sample libraries.

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

The sample RNA input can range from 25 ng to 1 µg.

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

The Ion AmpliSeq Mouse TCR Beta SR Assay, RNA is compatible with a vast array of research sample types, including FFPE tissue, fresh-frozen (FF) tissue, sorted T cells, whole blood, periperal blood leukocytes (PBLs), and periperal blood mononuclear cells (PBMCs).

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

The Ion AmpliSeq Mouse TCR Beta SR Assay, RNA (Cat. No. A45489) is a robust, targeted next-generation sequencing (NGS) assay designed to accurately identify and measure the clonal expansion of T lymphocytes by targeting the complementarity-determining region 3 (CDR3) of the T-cell receptor (TCR) gene locus from total RNA input.

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