Introduction:
In the dynamic landscape of DNA analysis, staying at the forefront of technology is paramount. VERSA Series innovative automation platform coupled with Promega’s PowerPlex Fusion 6C Amplification Kit, represents an innovative dive in genetic profiling. This powerful combination not only expedites the forensic DNA analysis process but also ensures precision, reliability, and scalability.
The PowerPlex Fusion 6C Amplification Kit:
Multiplex amplification and profiling of short tandem repeat (STR) loci provide a swift means of characterizing forensic samples for human identification. Various multiplex kits, such as the PowerPlex® Fusion 6C Systems provided by Promega Corporation, are available for consideration and procurement by DNA testing laboratories. In this context, we automate comprehensive protocols for both full-volume and half-volume amplification using extracted DNA for these multiplex kits.
Promega’s PowerPlex® Fusion 6C System Amplification Kit is a 27-locus multiplex for human identification, a solution designed for multiplex PCR applications. Specifically developed for the amplification of short tandem repeat (STR) markers, this kit empowers laboratories with the ability to simultaneously analyze multiple genetic loci. The inclusion of six fluorescent dyes enhances the discrimination power, providing comprehensive DNA profiles crucial for forensic investigations and paternity testing. This six-color system allows co-amplification and fluorescent detection of the 20 autosomal loci in the expanded CODIS core loci.
VERSA Instrument Preparation:
The VERSA Series automation platform reflects the ongoing advancements in laboratory automation. Its advanced features and adaptability streamline workflows, offering laboratories unparalleled efficiency and accuracy. When integrated with Promega’s PowerPlex® Fusion 6C Systems, the VERSA platform transforms DNA analysis, automating high-throughput processes and delivering results with unmatched reliability.
Promega PowerPlex® Fusion 6C protocol
Table 1: Amplification of Extracted DNA in a 25μl Reaction Volume
PCR Amplification Mix Component | Volume Per Reaction | Number of Reactions | Final Volume (48 samples) | Time calculated (48 samples) |
Water, Amplification Grade
| 5μl | 48 | 250μl
| 5 min |
PowerPlex ® Fusion 6C 5X Master Mix | 5μl | 48 | 250μl | |
PowerPlex ® Fusion 6C 5X Primer Pair Mix | 5μl | 48 | 250μl | |
Template DNA (1ng) | 10μl | 48 | 500μl | |
Total reaction volume | 25μl | 1250μl | ||
Dispense the PCR mix into each well of the PCR plate | 25μl | 6 min | ||
ProFlex™ PCR System or GeneAmp® PCR System 9700 | Thermocycler Off deck | Manual |
Instruments needed:
- VERSA Series
- GeneAmp® PCR System 9700 with a gold-plated silver or silver sample block, Veriti® 96-Well Thermal Cycler or ProFlex™ PCR System (Applied Biosystems)
- Centrifuge compatible with 96-well plates or reaction tubes
- MicroAmp® optical 96-well reaction plate or 0.2ml MicroAmp® reaction tubes (Applied Biosystems)
- Aerosol-resistant pipette tips
Amplification Setup
- At the first use, thaw the PowerPlex® Fusion 6C 5X Master Mix, PowerPlex® Fusion 6C 5X Primer Pair Mix and Water, Amplification Grade, completely. After the first use, store the reagents at +2°C to +10°C.
- Determine the number of reactions to be set up. This should include positive and negative control reactions.
- Use a clean plate for reaction assembly, and label appropriately. Alternatively, determine the number of clean, 0.2ml reaction tubes required, and label appropriately.
- Add the final volume of each reagent listed in Table 1 to a clean tube.
- Set the protocol in VERSA software and RUN.
Deck layout of VERSA
Fig. 1 VERSA liquid handling workstation as it pertains to the Promega PowerPlex® Fusion 6C 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 deck layout is optimized to minimize the mobile distances of the pipette head, thus saving off a minute or two of runtime.
The VERSA deck layout includes the following modules
- 8 channel pipette head with single channel functionality
- Gripper
- HEPA Filtered Hood with UV fluorescent light
- Slot for 20 µl, 200 µl, 500 µl and 1000 µl pipette tip boxes.
- Reagent block cooler for PowerPlex® Fusion 6C Kit components. 39 positions cooler: For storing buffers, reagents and enzymes at 4°C.
- Cooler/Heater Flat Plate (4°C-90°C)
- 8-well special reservoir
VERSA Software
Fig. 2 The VERSA1100 workstation is powered by the user-friendly VERSAware Plus 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 Promega PowerPlex® Fusion 6C 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.
Conclusion:
The combination of Promega’s PowerPlex Fusion 6C Amplification Kit with the VERSA Series automation platform marks an exciting step forward in DNA analysis techniques such as Capillary Electrophoresis and Next Generation Sequencing. Laboratories using this setup can benefit from greater efficiency and accuracy, helping them keep up with the growing demands of forensic science and overcome the backlog. As technology advances, the connection between advanced molecular biology tools and automation platforms is likely to reshape genetic profiling, highlighting the importance of automation in the future of DNA analysis.