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Making Forensic DNA Analysis smooth: Automating the Quantifiler® Trio DNA Quantification Kit on VERSA 1100

When it comes to securing forensic DNA analysis, something that’s not just good but absolute impressive is needed. Well, say hello to the VERSA 1100 automation workstation, a game-changer in the field, now automating the Quantifiler Trio DNA Quantification kit to make manual protocols smoother and faster. This synergy of cutting-edge technology brings forth unparalleled benefits, redefining efficiency and reliability in forensic laboratories.

Understanding the Quantifiler Trio DNA Quantification Kit

The Quantifiler® Trio DNA Quantification Kit is designed specifically for forensic DNA analysis. It provides a comprehensive solution for simultaneously quantifying three key genetic targets: human DNA, male DNA, and total DNA, enabling a deeper understanding of the sample composition and aiding in the interpretation of complex forensic samples. Once DNA is isolated from biological samples, measuring amplifiable human DNA becomes crucial for DNA analysis. The Quantifiler® Trio kit offers an easy method to accurately gauge human and male DNA concentrations, even as low as 5 pg/μL. This blog layouts the automation preparation and processing of quantitation reactions using the Quantifiler® Trio kit.

Deck layout of VERSA

VERSA liquid handling workstation as it pertains to the Quantifiler® Trio DNA Quantification Kit.  This particular deck layout is designed for up to 96 samples.  It could easily be expanded well beyond this for multiple plate automation for high throughput protocols.  The total runtime for 48 samples is about 11 minutes.

The VERSA deck layout includes the following modules

  • 8-channel pipette head with gripper
  • HEPA Filtered Hood with UV fluorescent light for VERSA series
  • Slot for 20 µl, 200 µl and 1000 µl pipette tip boxes.
  • Reagent block cooler for Quantifiler Trio DNA Quantification Kit tubes. 39 positions cooler: For storing buffers, reagents and enzymes at 4°C. It contains 16 places for 0.2mL, 12 for 0.5mL, 5 for 1.5mL and 6 for 5-7mL.
  • Cooler/Heater Flat Plate (4°C-90°C)
  • 8-well special reservoir

VERSA Software

The VERSA1100 workstation is powered by the user-friendly VERSAware software, which features a deck layout that allows users to select plate types from a library of existing labware. Software scripts can be created or modified using a drag-and-drop feature from the library of action-based commands. These features make it easy for users to create and modify scripts as needed. However, for Quantifiler® Trio DNA Quantification Kit workflows that are already pre-programmed, no modifications are needed. The pre-programmed workflows are designed to work seamlessly with the VERSA1100 platform and can be easily accessed through the software interface.

Streamlined Workflow

The VERSA 1100 transforms the DNA quantification process into a seamless automated workflow. From sample preparation to quantification, this integration minimizes manual intervention, significantly reducing human error and maximizing throughput. It can be configured starting from 24-sample upto 96-well format depending on instrument channels (4 or 8-channel head) – actual throughput time will depend on the specific procedure and steps involved. Laboratories can now process higher sample volumes with consistent and reliable results, all within a fraction of the time.

StepPosition From-ToVolumeTime (48 samples)
Primer MixReagent cooler- Reagent cooler8 μL1 min
Reaction MixReagent cooler- Reagent cooler10 μL1 min
Dispense the Master Mix into tubesReagent cooler- Reagent cooler18 μL3.5 min
Dispense the PCR mix into each well of the PCR plateReagent cooler- PCR plate18 μL2.5 min
Add sample, standard, or control to the applicable wells or tubes (Tip changer)Sample Plate- PCR plate2 μL3 min
Centrifuge the plate at 3000 rpm for about 20 seconds and run the plate on 7500 Real-Time PCR instrumentOff Deck

Once the automation run concludes, the Standards and sample DNA tubes undergo manual vortexing, centrifugation, followed by determining the concentration of the sample DNA.

Results

This automated Quantifiler protocol yields accurate and reproducible DNA quantitation results.  As part of QC (quality control) tests, numerous automated Quantifiler runs compared side-by-side with manual Quantifiler methods. The corresponding measured DNA concentrations were within the margin of error for the Qubit fluorometer. Thus, we feel confident that automated DNA quantitation is equivalent to manual quantitation with respect to the accuracy of results.

Additionally, this automated approach significantly reduces the likelihood of errors present in manual procedures. While skillful laboratory technicians and analysts can execute the Quantifiler protocols with great accuracy, the repetitive nature of these protocols, when applied to numerous samples, might induce mental fatigue, potentially resulting in occasional operational errors. These repetitive tasks can effectively be conducted by automated liquid handling workstations instead.

Eventually, employing automation for DNA quantification allows laboratory researchers to allocate their time to other crucial and higher-value tasks. The considerable time saved through the utilization of VERSA series is substantial.

Different Kits can be automated on VERSA series:

Conclusion

The automation of the Quantifiler Trio DNA Quantification kit on the Aurora VERSA 1100 automated liquid handling workstation offers a solution for high-throughput quantification, reducing hands-on time and human error with a total runtime of only 11 minutes for 48 samples setup. VERSA liquid handling systems have the advantage of having a user friendly interface, with flexible options for modifying the number of libraries, the number of qPCR replicates, and the dilution factor to desired experimental design with a maximum of 96 samples per run. Reliable, optimized and fully walk away comprehensive solution for quantitative studies.

So, dive into the world of automation with the VERSA 1100 and watch your lab zoom past the competition in forensic DNA analysis!