Quick and accurate processing of buccal or blood samples gathered via swab and paper substrates is vital to the efficient operation of forensic laboratories. The GlobalFiler® Express Kit* is an STR multiplex assay that allows the direct amplification from reference samples including paper punches and swabs.
This article outlines the utilization of the FORMULATRIX® MANTIS® to dispense the GlobalFiler® Express Kit master mix to the amplification plate with treated buccal punches present.
Methods and materials
Before the addition of GlobalFiler® Express Kit reagents the 1.2 mm paper disks treated with sample were added to the number of wells required. The GlobalFiler® Express Kit reagents were processed according to the vendor’s instructions for 26 cycle treated paper direct amplification.
The master mix was prepared by combining the following volumes per sample:
- 6.0 μL primer set
- 6.0 μL master mix
- 3.0 μL low-TE buffer
As seen in Figure 1, the MANTIS® liquid handler was utilized to dispense 15 μL master mix to the MicroAmp® Optical 96-Well Reaction Plate. The subsequent amplification products were run on an Applied Biosystems 3500xl Genetic Analyzer and analyzed with GeneMapper® ID-X v1 .4 software**.
Figure 1. MANTIS® Liquid Handler. Image Credit: FORMULATRIX
Results
Figure 2. Profile generated from a treated buccal punch us-ing GlobalFiler® Express at 26 cycles. Image Credit: FORMULATRIX
As seen in Figure 2, of the 17 treated buccal punches tested, all 17 produced complete profiles using a 26 cycle PCR amplification. All amplification controls (two positive and two negative controls) also produced the expected results.
Conclusion
The combination of the FORMULATRIX® MANTIS® liquid handler and the GlobalFiler® Express PCR Amplification Kit produces an efficient system for creating profiles. The GlobalFiler® Express chemistry eliminates the requirement for extracting and quantifying DNA before amplification.
Controlled by intuitive software, the MANTIS® supplies accurate and robust automated dispensing of the GlobalFiler® Express Master Mix to the amplification plate with minimal dead volume.
A complete system for processing paper punches with complete chain of custody would be provided by the integration of an automated punching robot and a LIMS system.
In order to meet the automation needs of modern forensic laboratories, future studies will include dispensing GlobalFiler® Express PCR Amplification reagents with a newer Mantis featuring pressurized bottle dispensing.
References
* For Forensic or Paternity Use Only.
** For Research, Forensic, Paternity and Cell Line Authentication. Not for use in Diagnostic and Therapeutic applications. GlobalFiler Express is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.
About FORMULATRIX
FORMULATRIX® was established in 2002 to provide protein crystallization automation solutions. Since then, we've started developing other laboratory automation solutions including the next generation of liquid handlers using microfluidic technology.
Headquartered in Bedford, Massachusetts, we supply software and robotic automation solutions to leading pharmaceutical companies and academic research institutions around the world. Our team works tirelessly to provide the best products in the industry with support that is second to none.
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