Before preparing libraries, record information about your samples using an option available for your sequencing system:
The AmpliSeq for Illumina Focus Panel is a targeted resequencing assay for biomarker analysis of 52 genes with known relevance to solid tumors.
| Component | Number of Pools | Concentration | Number of Amplicons | Average Amplicon Length (bp) | Average Library Length (bp) |
| DNA | 1 | 5X | 269 | 107 | 247 |
| RNA | 1 | 5X | 284 | 93 | 233 |
Completing the AmpliSeq for Illumina Focus Panel protocol requires the panel and the following library prep, cDNA synthesis, and index kits. The AmpliSeq for Illumina Library PLUS Kit is available in 24-, 96-, and 384-reaction configurations.
The AmpliSeq for Illumina Focus protocol generates one DNA and one RNA library per sample. Samples can be DNA, RNA, or paired. Samples for this protocol are paired. A sample is a specimen of nucleic acid from one source, while a library is nucleic acids from a sample after preparation. A library is a unique DNA or RNA preparation that 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 Libraries | 48 Libraries (24 DNA, 24 RNA) |
192 Libraries (96 DNA, 96 RNA) |
768 Libraries (384 DNA, 384 RNA) |
| AmpliSeq for Illumina Focus Panel | 1 | 4 | 16 |
| AmpliSeq Library PLUS For Illumina | 2 24-reaction kits | 2 96-reaction kits | 2 384-reaction kits |
| AmpliSeq CD Set A (96 reaction, 96 indexes) | 1 | 2 | 8 |
| cDNA Synthesis | 1 | 1 | 4 |
Sequence AmpliSeq for Illumina Focus Panel libraries using the following systems and reagent kits.
| System | Reagent Kit | Samples per Run |
| MiniSeq System | MiniSeq Mid Output Reagent Kit | 24 |
| MiniSeq High Output Reagent Kit | 72 | |
| MiSeq System | MiSeq Reagent Micro Kit v2 | 12 |
| MiSeq Reagent Kit v2 | 40 | |
| MiSeq Reagent Kit v3 | 72 |
| System | Reagent Kit | Samples per Run |
| MiniSeq System | MiniSeq Mid Output Reagent Kit | 48 |
| MiSeq System | MiSeq Reagent Kit v2 | 96 |
| MiSeq Reagent Nano Kit v2 | 6 | |
| MiSeq Reagent Micro Kit v2 | 24 |
| System | Reagent Kit | DNA Samples per Run | RNA Samples per Run | Combined Samples per Run |
| MiniSeq System | MiniSeq Mid Output Reagent Kit | 16 | 16 | 32 |
| MiniSeq High Output Reagent Kit | 48 | 48 | 96 | |
| MiSeq System | MiSeq Reagent Micro Kit v2 | 8 | 8 | 16 |
| MiSeq Reagent Kit v2 | 30 | 30 | 60 | |
| MiSeq Reagent Kit v3 | 48 | 48 | 96 |