MiSeq FGx is a benchtop sequencer designed for forensic genetics applications. Together with the dedicated server incorporating ForenSeq Universal Analysis Software (UAS), it enables users to configure runs, sequence libraries, analyse profiles and generate reports through a workflow adapted to forensic genetics applications. The application and type of report depend on the kit and the version of the UAS module used.
Detailed description
Operating principle
The previously amplified libraries are loaded into the reagent cartridge and sequenced on MiSeq FGx using Illumina sequencing-by-synthesis (SBS) chemistry. The instrument guides loading and run configuration through a touchscreen. The independently installed server running the UAS software receives the data and performs the analysis corresponding to the assay: FASTQ generation, alignment, variant calling and reporting of results according to the available module.
Applications
Benefits
The UAS server manages projects and samples, analyses results and generates assay-specific outputs: CODIS reports for MainstAY, EMPOP reports for mtDNA and GEDmatch PRO reports for Kintelligence.
Key indicators
| MiSeq FGx parameter | Data specified by QIAGEN |
|---|---|
| Maximum read length | 2 × 300 bp, depending on the assay |
| Output for a 2 × 150 bp run | ≥5 Gb; 12.5 million read pairs passing filters |
| Quality at 2 × 150 bp | ≥80% of bases at Q30 |
| Overall accuracy and reproducibility | ≥99.66% and 99.7%, respectively |
| Dimensions and weight | 68.6 × 56.5 × 52.3 cm; 54.5 kg |
| Samples per run | 1–384, depending on the assay; actual capacity depends on the kit |
The performance figures are the generic specifications of the instrument. The capacity and coverage for each marker in each analysed sample depend on the programmed assays and their configuration.
The generated libraries can be sequenced using two cartridges with different capacities, providing considerable flexibility when configuring assays. The standard cartridge provides up to 12.5 million read pairs (25 million individual reads), while the Micro cartridge provides up to 5 million pairs.
Intended users
Forensic genetics and human identification laboratories seeking to incorporate massively parallel sequencing into their studies and centralise analysis on a dedicated server.
Considerations
The instrument, UAS server, sequencing reagents and library preparation kits have separate reference numbers. Before selecting an assay, its compatibility with the installed UAS version, cartridge type and required multiplexing must be confirmed.
Key aspects
Presentation details
| Product | Presentation | Reference |
|---|---|---|
| MiSeq FGx Sequencing System | Benchtop instrument | 15048976 |
| ForenSeq Universal Analysis Software | Server with UAS pre-installed, monitor, keyboard and mouse | 9003364 |
| MiSeq FGx Reagent Kit | Standard sequencing cartridge | 15066817 |
| MiSeq FGx Reagent Micro Kit | Micro sequencing cartridge | 20021681 |
