Before preparing libraries, record information about your samples using an option available for your sequencing system:
The AmpliSeq for Illumina Cancer Hotspot Panel v2 is a targeted resequencing assay for researching somatic mutations across the hotspot regions of 50 genes with known associations to cancer, as identified in the Catalogue of Somatic Mutations in Cancer (COSMIC) database.
| Number of Pools | Concentration | Number of Amplicons | Average Amplicon Length (bp) | Average Library Length (bp) |
|---|---|---|---|---|
| 1 | 5X | 207 | 106 | 246 |
Completing the AmpliSeq for Illumina Cancer Hotspot Panel v2 protocol requires the panel and the following library prep and index kit. The AmpliSeq for Illumina Library PLUS Kit is available in 24-, 96-, and 384-reaction configurations.
For each DNA sample, the AmpliSeq for Illumina Cancer Hotspot Panel v2 generates one DNA library. A sample is a specimen of nucleic acid from one source, while a library is nucleic acids from a sample after preparation. A library has the ends required for sequencing.
Preparing the appropriate number of samples requires the following number and combination of kits.
| Number of Samples | 24 Samples | 96 Samples | 384 Samples |
|---|---|---|---|
| Number of Libraries | 24 Libraries | 96 Libraries | 384 Libraries |
| AmpliSeq for Illumina Cancer Hotspot Panel v2 | 1 | 4 | 16 |
| AmpliSeq Library PLUS For Illumina | 1 24-reaction kit | 1 96-reaction kit | 1 384-reaction kit |
| AmpliSeq CD Set A (96 reaction, 96 indexes) | 1 | 1 | 4 |
| cDNA Synthesis | Not applicable | Not applicable | Not applicable |
Sequence AmpliSeq for Illumina Cancer Hotspot Panel v2 libraries using the following systems and reagent kits.
| System | Reagent Kit | Samples per Run |
|---|---|---|
| MiniSeq System | MiniSeq Mid Output Reagent Kit | 32 |
| MiniSeq High Output Reagent Kit | 96 | |
| MiSeq System | MiSeq Reagent Nano Kit v2 | 4 |
| MiSeq Reagent Micro Kit v2 | 16 | |
| MiSeq Reagent Kit v2 | 60 | |
| MiSeq Reagent Kit v3 | 96 |