Qiagen, 15048976, MiSeq FGx Sequencing System

MiSeq FGx Sequencing System is a desktop NGS instrument with integrated software for run setup, results interpretation and reporting

Features

- High-performance benchtop sequencer with intuitive touch‑screen and load-and-go reagents
- Advanced capacity to process challenging DNA samples
- Interrogates hundreds of markers in a single workflow
- Optimized flexibility for human identification applications

Product Details

The MiSeq FGx Sequencing System is the first and only next-generation sequencing (NGS) instrument designed, developed and validated for forensic genomics. Combining proven data quality with ease of use, the system is key to a unique, single-platform solution built on gold-standard NGS technology. Prepare and sequence libraries and analyze data in a single workflow designed to scale for a growing portfolio of applications, including new tools like Forensic Investigative Genetic Genealogy (FIGG). Dedicated reagent kits and matched analysis software empower answers for all human identification investigations.

Performance

Feature: Details
Throughput: 1–384 samples per run, depending on assay
Maximum read length: 2 x 300 bp, depending on assay
Output (2 x 150 bp run): ≥ 5 Gb
Reads passing filters: 25 million
Q30 score (at read length of 2 x 150 bp): ≥ 80%
Total overall accuracy: ≥ 99.66%
Total overall reproducibility: ≥ 99.7%
Biomolecules that can be sequenced: gDNA, RNA, mtDNA
Modes: Forensic (FGx) and Research Use Only (RUO)
Power requirements: 100–240 V AC at 50/60Hz, 10A, 400 W
RFID radio frequency: 13.56 MHz
RFID power: 100 mW
Dimensions: 68.6 cm × 56.5 cm × 52.3 cm (27 in × 22.2 in × 20.6 in)
Weight: 54.5 kg (120 lbs)

The MiSeq FGx System is integral to a validated workflow designed specifically for forensic laboratories. The system is fully supported with performance qualifications and available maintenance that align to requirements of a forensic laboratory. Building on the speed and accuracy of Illumina sequencing-by-synthesis (SBS) chemistry, the MiSeq FGx System brings the unprecedented focus of forensic genomics to NGS, granting criminal justice a powerful ally.

Superior analysis of complex DNA mixtures and degraded DNA

Integrated sequencing workflow consolidates library prep, sequencing, and analysis

Advanced capability with adjustable read lengths, dual run modes and two flow cells

The MiSeq FGx System is the first and only instrument to interrogate up to 96 combined SNP and STR libraries in a single run. The MiSeq FGx System preserves precious sample while demonstrating robust performance, developing more thorough, detailed DNA profiles from a wide range of sample types, biological molecules, and across the quality spectrum of high-quality genomic DNA (gDNA) to degraded, mixed and limited samples.

Two run modes promote a range of forensic genomics applications while retaining the flexibility to work in a research environment. Easily switch between two sequencing kits to tune output for the current application or sample batch size. Built on industry-leading Illumina SBS chemistry, the MiSeq FGx System meets strict performance requirements for comprehensive and conclusive results. Virtually eliminate context-specific errors, even within repetitive sequence regions or homopolymers and answer a wider range of questions in one targeted assay with multiplexing capability. An intuitive touchscreen interface simplifies instrument operation and the system monitors quality and performance metrics during a run. After sequencing, analysis starts automatically and completes in under one hour.

Principle

Compared to size-based analysis of short tandem repeats (STRs) using capillary electrophoresis (CE), NGS detects the full spectrum of genetic variation in a DNA sample with a faster time to results than other technologies. Inherently sensitive, ForenSeq NGS technology goes beyond fragment size to identify underlying sequence variation. Comprehensive results are delivered quickly and clearly, displaying base-by-base sequences for easy interpretation with utmost confidence. Building on STR capabilities, NGS brings other advantages to modern forensic genomics, including analysis of nuclear single nucleotide polymorphisms (SNPs), mitochondrial DNA (mtDNA), messenger RNA (mRNA) and epigenetic markers, bolstering cases that require human identification while eliminating the need to weigh technical limitations against potentially informative data. Achieve superior results in a single sequencing run.

The MiSeq FGx System achieves exceptional data quality by employing a proprietary, reversible terminator‑based method that detects single bases as they are incorporated into massively parallel DNA strands. Fluorescent terminator dyes are imaged as each deoxynucleotide triphosphate (dNTP) is added and then cleaved to allow incorporation of the next base. With all four reversible, terminator-bound dNTPs present at each sequencing cycle, natural competition among bases minimizes incorporation bias. The software makes base calls directly from signal intensity measurements during each incorporation cycle, reducing raw error rates compared to other technologies. The result is highly accurate, base-by-base sequencing that minimizes sequence context‑specific errors, even within repetitive sequence regions or homopolymers.

Procedure

Libraries sequenced on the MiSeq FGx System follow the same core NGS workflow: library prep, sequencing and analysis. The first step uses a library prep kit to add primers to genomic DNA (gDNA), mtDNA or mRNA extracted from forensic samples for simultaneous capture and amplification of hundreds of target regions, generating dual-indexed libraries ready for sequencing. After library prep, load-and-go reagents streamline run setup. Simply thaw the prefilled reagent cartridge, add libraries to the cartridge, and insert the cartridge into the instrument. The system software accepts run parameters and starts sequencing with the push of a button.

Applications

The MiSeq FGx System enables capabilities across all biological molecules and a growing number of applications, including both short- and long-range kinship analysis. Adjustable read lengths, flow cell options and choice of run modes allow unprecedented flexibility for matching data output to an ever-increasing range of human identification needs. The MiSeq FGx System delivers an enhanced capacity to analyze degraded DNA, low-quantity DNA, complex DNA mixtures and other challenging samples that can complicate or derail an investigation. Small amplicon sizes are well suited for interrogation of degraded DNA, while large numbers of markers—including many that are highly polymorphic—improve system ability to discern low-level minor components that CE might not detect. Additionally, one sequencing run interrogates hundreds of forensically relevant genetic markers, eliminating the need to choose between fragment length‑based STR kits or otherwise make trade-offs to accommodate challenging samples.