Automate On-Chip DNA Library Prep Without Bulky Instruments:

On-chip fluid handling system with the footprint of a hand-held device

Problem

Large instruments with complex tubing are often required to automate DNA library preparation for NGS and other genetic analyses. This makes the system complex, cumbersome, and expensive. Without frequent flushing, transferring DNA samples to different reaction sites via tubes can create contamination risks. Contamination during DNA library prep can decrease sequencing yield and create sequencing problems. Finally, most laboratories have limited space for instruments, making device footprint size an essential concern for labs in life science and biotechnology industries.

Solution

Enplas’s rotary membrane valve and pump system (RMVP) can automate your DNA library prep with a significantly reduced instrument size. A hand-held size device operates multiplex fluid handling, which simplifies automation without risk of contamination. The semi-closed environment reduces tubing and prevents the risk of contamination. Additionally, a microfluidic chip contains accurately sized reservoirs and channels to meter, mix, sort, or incubate various samples and reagents.

On-Chip DNA Library End Prep

In the following video, the RMVP achieves automation of DNA sample prep through the pulling, pushing, and mixing of reagents using accurate metering chambers. The RMVP completes DNA library prep entirely within the semi-contained system. Bead solutions are metered via the same on-chip functions and pushed to on-chip PCR chambers for thermal cycling. The valve pusher also carries out magnetic bead mobilization and immobilization using integrated magnets.

The RMVP achieves automation of DNA library prep, including PCR, size selection, and bead isolation by magnets, all on one chip.

Customizing Microfluidic Channels

Creating the appropriate vessel for the RMVP to conduct all the above reactions is critical. This is done by customizing the microfluidic chip channels. Accurately sized reservoirs and channels can meter, mix, sort, or become incubation sites for various samples and reagents. In addition, precision injection molding is required for microfluidic chips to create exact copies of highly precise and sensitive chip features–that require micrometer accuracy during production to create the exact required reactions in millions of units.

Benefits

The RMVP system is versatile and can be customized freely to meet your unique application needs. In this example, the Enplas RMVP system achieved the automation of the end prep process of a DNA library prep, which included PCR, size selection, and beads isolation by magnets, all on one chip, within a handheld-sized device.
The RMVP controlled fluids to move back and forth, mix, and meter the exact amounts for reactions on-chip without an external pump. Twelve reagents were used in this example, but the RMVP can control more than 20 different fluids without contamination.
The RMVP system eliminates the need for instrument tubing, which makes the system larger, more complex, and prone to error or contamination.
All reactions are conducted in a semi-closed environment, reducing amplicon cross-contamination risks.
The RMVP runs with less energy consumption and without excessive heat emission, in contrast to larger instruments that require multiple motors for various valves and pumps.

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Specifications

Reagents used in the video

PCR enrichment master mix
PCR enrichment primer
Size selection bead
Uracil depletion enzyme
1x TE
80% ethanol
Fragmented DNA
End prep enzyme
End prep reaction buffer
Adapter
Ligation master mix
Ligation enhancer

Useable Materials for the Plastic Chip

Example Applications

DNA/RNA purification
Sample prep / Library prep
PCR
NGS
Other molecular tests and research

Request a Demo of the RMVP

There are various ways to utilize the push-pull function to achieve the required fluid motions and mixing needs for your specific application. The RMVP is best experienced yourself. To request a demo, click here.