D4 FAQs
D4 - Frequently Asked Questions
What is the purpose of the D4 Digital Sample Preparation System?
The D4 is designed for automated library preparation, which includes tasks such as enzymatic fragmentation, end repair, adapter ligation, PCR, magnetic bead purification, fragment size selection, nucleic acid quantification, and DNB creation. Users can input DNA and/or RNA samples into the system and receive the final product, which could be double-stranded DNA or DNB. The entire process is automated, and the system also provides the product concentration and the recommended input amount for sequencing.
What is the sample throughput of the D4?
The D4 processes one sample preparation cartridge at a time, and each cartridge contains 4 lanes. Typically, it supports up to 4 samples per run.
How long does the process take (turn-around time) on the D4?
After adding sealing fluid, library reagents, and samples (a process that takes approximately 15 minutes), the sample preparation cartridge is inserted into the D4, which begins the automated library preparation process. The turn-around time (TAT) depends on the library protocol used. For example, using the D4RS Enzymatic DNA Library Preparation Kit V2.0 (940-001198-00), which includes steps like enzymatic fragmentation and DNB creation, the TAT is around 3 hours, including automated quantification. Generally, the TAT for the D4 is comparable to or shorter than manual library preparation.
Does the D4 have a built-in quantification function?
Yes, the D4 features a built-in quantification module that automates the process of quantifying double-stranded DNA libraries or DNBs using fluorescent dyes. It displays the product concentration and the recommended input amount for sequencing on the result page.
What manual operations are required for the D4?
For the standard library preparation kit, users need to manually add sealing fluid, library reagents, and samples into the sample preparation cartridge, a process that takes under 15 minutes. After the cartridge is inserted into the instrument, the D4 automatically completes the rest of the library preparation steps. The result page shows the quantitative results for each lane. Once the process is finished, users remove the cartridge, extract the product, and discard the sealing fluid. Product pooling can be done as needed for sequencing.
How does the D4 prevent aerosols and cross-contamination?
The D4 is built with a fully closed system that ensures no aerosols are produced during the library preparation process. All droplets remain sealed under fluid, and all waste generated is contained within the preparation cartridge. This keeps all reactions isolated from the external environment, preventing contamination. The system has been validated to eliminate cross-contamination across various applications, including mNGS, tNGS, and small cancer panels, ensuring zero-contamination library preparation.
