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 video to the left, the RMVP automates DNA sample prep within a semi-closed system by mixing reagents through simple pulling and pushing functionalities. Solutions are precisely metered via accurately manufactured on-chip chambers, then pushed to on-chip PCR chambers for thermal cycling. When magnetic beads are necessary, magnets can also be integrated into the RMVP system to control mobilization and immobilization of the beads.
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.
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:
- COC
- COP
Example Applications:
- DNA/RNA purification
- Sample prep / Library prep
- PCR
- NGS
- Other molecular tests and research
