Ion AmpliSeq™ 样品 ID 检测组合

A DNA fragment library is constructed from whole genomic DNA and is commonly used for whole genome resequencing or de novo sequencing. Briefly, the whole genomic DNA is fragmented or sheared, ligated with Ion-specific adapter sequences, and then size-selected for the library fragments of the desired length.

Amplicon libraries are constructed from PCR-amplified DNA fragments and are used for targeted sequencing (e.g., investigating variants at known genomic locations). There are two types of amplicon libraries, short and long.

A short amplicon library contains DNA fragments (targets) with lengths that are compatible with the Ion template preparation kits without any further shearing or fragmentation during library preparation. Additionally, no size-selection step is required, as the amplicons are already within the desired size range.

A long amplicon library contains DNA fragments (targets) with lengths that are longer than those compatible with the Ion template kits and requires further shearing or fragmentation during library preparation. The library preparation protocol for long amplicons is similar to fragment libraries.

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

The Ion AmpliSeq Sample ID Panel (Cat. No. 4479790) is a versatile, cost effective, and easy-to-use human SNP genotyping panel comprising 9 specially designed primer pairs that can be added to the target amplification reaction to generate a unique ID during post-sequencing analysis of research samples using the on AmpliSeq Sample ID Panel plug-in. This panel is useful for tracking tumor/normal paired samples, samples from the same individual in longitudinal studies, and multi-tissue or multi-tumor samples from the same individual.

The Ion AmpliSeq Sample ID Panel can be used in conjunction with all Ion AmpliSeq Ready-to-Use, Custom, and Community panels.

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

If you've reviewed your design and are not satisfied with the results, please click on the “Not happy with this design? Let us help” link to have an Ion AmpliSeq team member contact you about additional options for your design.

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

The ~70 bp or ~90 bp peak is likely standard or barcoded adapter dimers, respectively. Adapter dimers may form during the adapter ligation step and are usually removed during the size selection process. The adapter dimers will amplify on the Ion Torrent Ion Sphere particles during template preparation and decrease the overall throughput of usable sequencing reads; thus, we highly recommend removing the adapter dimers by performing an additional clean-up step prior to template preparation.

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

In addition to input RNA quality and accurate quantification, the clean-up and size selection steps are critical to generating a successful RNA-Seq library.

- Be sure to mix the nucleic acid binding beads well before dispensing, and follow the workflow and incubation times as closely as possible.
- Use fresh ethanol and pre-wet pipette tips prior to transferring ethanol, as the volume is critical for size selection.
- Remove residual ethanol before elution using a small-volume pipette. Do not over-dry or under-dry the beads.

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