Frequently asked questions for AmpliSeq for Illumina On Demand panels

AmpliSeq for Illumina On-Demand panels provide a choice of > 5000 pretested genes with known content relevant for inherited disease research, including hereditary cancer, primary immunodeficiency, hearing loss, and muscular dystrophy.

• Can the On-Demand panel be mixed with another custom panel?

  • No, each custom panel (On-Demand, Gene, Hotspot) is its own, distinct panel.

• How many PCR cycles should be used to amplify the custom targets?

  • Use the number of Primer Pairs as listed on the tube to inform the number of PCR cycles to use. The reference guide provides recommended PCR cycles based on primer pair/amplicon number.

• What are the minimum and maximum content (target) sizes?

  • Amplicons: 24-15,000 amplicons

  • Genes: 1-500 genes

• What is the amplicon length?

  • All On-Demand panels have a maximum amplicon size of 275 bp.

• Which genome builds are supported?

  • Human hg19 only.

• What gene format is needed when adding targets in DesignStudio?

  • HGNC format.

• How are the amplicons tested?

  • Genes have been tested via sequencing on the NextSeq 550 with a 2x151 run.

  • Not all gene combinations have been tested.

• What are the specifications for performance?

  • As this is a custom AmpliSeq design, Illumina does not provide specifications for performance.

For any feedback or questions regarding this article (Illumina Knowledge Article #1048), contact Illumina Technical Support [e-mail].