Man-machine war (bottom): NGS sequence capture big competition

Blank in the second generation sequencing (NGS) process, the construction of gene libraries and target sequence capture and enrichment is the bottleneck restricting the sequencing process. In the last issue, we introduced an automation solution for building a database. Here we provide an automated solution for target sequence capture enrichment.

Blanks In recent years, second-generation sequencing has gradually evolved toward increasing throughput and reducing costs. To save analysis time and find a cost-effective approach, Roche Diagnostics developed the SeqCapEZ Capture Kit, which allows users to enrich the region of interest or the entire exome. But second-generation sequencing is a very time-consuming and labor-intensive task, often taking four days of professional technicians to complete. Moreover, some important considerations in the sequencing process also require the technician to maintain a high degree of concentration throughout the operation, such as capturing target sequences, heating kit samples, and controlling incubation time.

Blanks are subject to these limitations, and automating sequence capture is a requirement of most laboratories. Roche Diagnostics in France collaborates with PrimaDiag, a specialist automation platform manufacturer, to provide an automated solution for Roche NimbleGen exon capture sequencing products – the ACSIA NGS ^capture automated capture system. The ACSIA NGS ^capture system is available for all sequencing products from the Roche NimbleGen SeqCap Enrichment System.

The blank ACSIA NGS ^capture automated capture system uses PrimaDiag's innovative patented APIM to automatically process the beads, control the temperature to 0.1 °C, and the oscillation speed is adjustable from 200 to 3000 rpm. During the magnetic bead capture and washing phase, the heated wash solution is automatically completed. Wash the magnetic beads, incubate and shake the operation.

The blank ACSIA NGS ^capture system is flexible and easy to operate, allowing users to define the number of samples as needed. All liquid parameters are carefully optimized to minimize the actual reagent consumption used to build the library, and the optimized operating procedure allows the entire library to be completed with a minimum number of tips, saving significant reagent and consumable costs. For example, using Roche's SeqCap EZ kit for 48 reactions, the ACSIA NGS ^capture system requires only 3 cartridges, while manual completion requires 9 cartridges, which is three times the number of automated operations .

Blank Roche NimbleGen sequence capture kit has been carried out on ACSIA NGS ^capture system verification, do the steps to optimize, pre-running program, so using ^ACSIA NG Scapture automation platform to capture a sequence of interest, only need to open the default method, Enter the number of samples and click OK.

The blanks compared the effects of ACSIA and artificial capture. The sequencing results showed that the coverage depth of the 18 genes captured by the two methods was similar, and the ACSIA automated (blue) captured gene sequencing coverage was even slightly better than manual operation (red). See Figure 1.

The blank ACSIA NGS ^capture automatic capture system, flexible and simple operation, high-quality capture effect, and truly complete the NGS experiment.