Around the world, forensic DNA labs are being asked to do more with less. This is why the Applied Biosystems GlobalFiler IQC, GlobalFiler, and GlobalFiler Express PCR amplification kits combine reduced amplification time with exceptional discrimination power, even on the most challenging sample types.
As global forensic DNA databases rapidly expand, so does the need for more discriminating STR multiplexes that can maximize loci overlap. That’s why GlobalFiler kits incorporate the most commonly used loci—all in a single multiplex, 6-dye configuration kit. GlobalFiler kits contain all markers recommended for inclusion by the CODIS Core Loci Working Group and those markers commonly used in Europe. The multiplex selection of markers reduces the risk of adventitious matches while enabling more effective cross-border data sharing. And the kits are backed by training, service, and support from Thermo Fisher Scientific.
The GlobalFiler kit is also available with an Internal Quality Control system, or IQC, as part of a fully integrated and validated forensic workflow. The IQC system consists of two synthetic sequences with specific primers for each of the targets (IQCS and IQCL) and provides positive confirmation of sample amplification, but also indicates adverse conditions with compromised amplification, such as the presence of PCR inhibitors. The IQC system provides additional confidence in genotyping results and can help forensic scientists distinguish, for example, between inhibited and degraded DNA samples.
GlobalFiler kits are manufactured at our location in Warrington, UK, at a facility that meets the guidelines for ISO 18385 certification. We have made significant investments across all aspects of production to minimize human DNA contamination. The result: powerful forensic DNA-grade solutions that enable you to provide answers with certainty and confidence.
This breakthrough chemistry enables up to 5 times faster amplification time than previous-generation kits:
Typical workflow for up to 48 samples using Applied Biosystems direct PCR amplification kits, genetic analyzers and expert system analysis software.
The GlobalFiler Kit multiplex configuration includes all 24 loci with only 1 locus partially exceeding 400 base pairs. 10 mini-STR loci lie completely below 220 base pairs for enhanced performance on degraded samples, and all gender-specific markers are located in the green VIC channel for convenience of interpretation. The IQCS and IQCL markers are only present in GlobalFiler IQC kit.
The GlobalFiler Kit allelic ladder includes 343 alleles featuring expanded marker ranges for many loci while maintaining interlocus spacing. The expanded panel and virtual bins (589 total alleles) minimize OL allele calls and facilitate more accurate and efficient automated genotyping.
As global forensic DNA databases rapidly expand, so does the need for more discriminating STR multiplexes that can maximize loci overlap. That’s why GlobalFiler kits incorporate as many commonly used loci as possible—all in a single multiplex.
“Improving quality control processes and performance checks is a continuous challenge in forensics, more particularly for rapid, high-throughput, and reliable generation of DNA profiles from casework samples. The Internal Quality Control (IQC) system combined into GlobalFiler kits generates a powerful and controlled STR multiplex PCR assay for better and faster decision making and data interpretation.”
—Dr Sylvain Hubac
Research and innovation manager
High throughput DNA unit for casework samples
Forensic Science laboratory of the French Gendarmerie, France
Pablo Martín, Lourdes Fernández de Simón, Gracia Luque, María José Farfán, Antonio Alonso
Published online: July 15, 2014
For Forensic or Paternity Use Only.
For the past 16 years, Thermo Fisher Scientific has led the way in the evolution of STR technology. And today, GlobalFiler kits utilize breakthrough 6-dye chemistry to enable optimal performance and unprecedented discrimination.
Using a 6-dye configuration enables optimal performance, efficiency, data recovery and genotyping accuracy.
|GlobalFiler kit||GlobalFiler IQC kit||GlobalFiler Express kit|
|High discrimination power||24-marker multiplex assay including 3 gender markers and the highly discriminatory SE33 locus|
|Number of dyes||6|
|Mini-STR (<220 bp)||10|
|Gender markers||Y-indel, amelogenin, and DYS391|
|IQC markers||No||Yes: for distinguishing inhibited and degraded sample; positive control for PCR amplification||No|
|Identical primer sequences||✓||✓ (except for the IQC)||✓|
|Probability of identity (PI) value||African American: 6.8 x 10-27|
|US Caucasian: 3.71 x 10-26|
|US Hispanic: 3.09 x 10-26|
|Asian: 3.24 x 10-24|
|Required ESS markers||✓||✓||✓|
|Required CODIS markers||✓||✓||✓|
|NDIS* approved||✓||In progress||✓|
|DNAinput||15 μL/1 ng target||Treated or untrested paper: 1.2 punch Swab: 3 μL (of 400 L)|
Applied Biosystem Prep-n-Go Buffer
|Final PCR volume||25 μL||15 μL|
|Technical note supporting |
|Supported sample types||Optimized chemistry for challenging samples types: touched, inhibited, or degraded samples||Verified with multiple sample collection devices such as treated paper, untreated paper, and swabs; designed to work with the most commonly used substrates|
Dr. Antonio Alonso discusses how forensic laboratories can make the transition to 6-dye chemistries in order to gain workflow efficiency and reduce operating costs.
Recovering maximum information from degraded samples
In order to enhance information recovery from challenging bone samples, a recent degradation study was conducted by the University of North Texas Health Science Center, Department of Forensic and Investigative Genetics, US.
Look no further than our Human Identification Professional Services (HPS) team. Since 2007, we have completed over 400 successful validation projects worldwide with a team of more than 20 technical support specialists, each averaging 8 years of real-world forensic experience, providing customers with in-depth training and support on our instruments, chemistries, and software.
Q: Why do the Applied Biosystems GlobalFiler Kits utilize a 6-dye chemistry configuration that requires use of a 3500 or upgraded 3130/3730 Genetic Analyzer?
A: Squeezing all the additional loci into a 5-dye configuration would result in multiple tradeoffs, including:
Tradeoff #1: Multiple loci in the high molecular weight size range (>400 bp), increasing drop out with degraded and challenging samples.
Tradeoff #2: Several loci extending almost to 500 bp, which may cause issues with sizing and resolution, i.e. alleles labeled “OL” or failure to resolve microvariant alleles.
Tradeoff #3: Fewer miniSTRs. The GlobalFiler Kits include 10 STRs completely under 220 bp! This would not be possible in a 5-dye configuration.
Tradeoff #4: Insufficient spacing between adjacent markers; more difficult to expand marker ranges to minimize “OL” allele calls.
Tradeoff #5: Many loci would require redesigned primer sequences, leading to less concordance with data generated using the original primer sequences and with the vast majority of profiles in worldwide databases.
The bottom line is that using a 6-dye configuration enables optimal performance, efficiency, data recovery, and genotyping accuracy. Yes, this requires a 3500 or instrument upgrade, but isn’t this better than a chemistry downgrade?
Q: Why have you developed separate kits for casework and single source sample applications? Doesn’t this increase the validation burden on laboratories?
A: Our investigations indicate that to deliver the best direct amplification profile quality and efficiency for single source samples and the ultimate performance and specificity for casework analysis, the development of two application specific chemistries offers the best solution. To achieve the highest quality direct amplification results from a wide variety of single-source sample type/substrate combinations requires a buffer formulation that could result in compromised specificity for casework samples which we consider to be unacceptable. The validation burden may be slightly higher than for laboratories choosing a single kit and single reaction setup and/or amplification protocol for both casework and single-source samples; however, a little extra time spent on validation up front enables greater efficiency and performance for all samples types during routine operations which will mitigate any extra effort required during validation. In addition, the availability of our HPS (HID Professional Services) team can help shoulder the validation burden for the laboratory and thus helps reduce the impact of validation on precious laboratory time and resources.
For those laboratories that wish to utilize one kit for all sample types, we recommend the use of the GlobalFiler kit to prioritize the performance of casework samples over the less demanding single-source samples. We have demonstrated the feasibility of using the GlobalFiler kit in direct amplification mode.
Q: Why are there two Y chromosome loci when only one is needed to confirm a null allele in Amelogenin?
A: The DYS391 marker was listed as required in the expanded list of CODIS loci proposed by the US Federal Bureau of Investigation in January, 2012. To maximize spacing within the multiplex, we chose to locate DYS391 into the higher size range, reserving the smaller size range for the informative, autosomal loci. This does, however, render the DYS391 marker vulnerable to drop out in degraded samples. To address this, an extra gender determination marker in the form of a Y indel was added in the available space to the left of Amelogenin. This locus provides additional redundancy to Amelogenin, allowing confirmation of sample gender in severely degraded samples.
Q. Why did we not include rapidly mutating YSTR markers in the autosomal kit?
This adds complexity in data analysis. If it is confirmed through the Y marker that a male is present in the sample, then it is prudent to follow YSTR analysis. Moreover, mixture deconvolution is more complicated by the additional contributors present.
Q. What is the discriminatory power of the GlobalFiler kits?
Discrimination power comes into play with highly degraded or challenging samples that may yield only partial profiles. This is where the mini-STRs in the GlobalFiler kit come into their own with 10 loci under 220 BP and PI of loci <220 bp: 9.2 x 10-12 (increasing discrimination power by up to 9 orders of magnitude relative to older kits).
Q: Is there PCR cycle flexibility in using GlobalFiler kits?
In situations where you may observe data oversaturation, the cycle number can be reduced and a technical note is available on the same.
Q. What is ISO18385 and are the Globalfiler kits compliant?
ISO 18385 seeks to minimize human DNA contamination and GlobalFiler/Globalfiler Express kits are both compliant. Learn more ›
Q: Are the GlobalFiler Kit primer sequences compatible with previous AmpFlSTR kits?
A: Most loci contain the same core and SNP-specific primer sequences as found in the Identifiler series and NGM SElect Kits. To optimize position and spacing within the multiplex, the reverse primers were changed for the TPOX locus, compared to the one used in the Identifiler Kits, and DYS391, compared to the one used in the Yfiler Kit. Additional SNP-specific primers were added for the following loci to help minimize the occurrence of false homozygosity: D3S1358, vWA, D18S51, D19S433, TH01, FGA, D5S818, and SE33.
Q: What is the latest version of Data Collection software for the 3500 series instruments and what new features does it offer?
A: 3500 Data Collection Software v3.1 is the latest version of software for the 3500 and offers the following features/benefits
* On-instrument supported limits are the lower of: 14 days, stated number of injections, or the expiry date.
Q: Do I need to upgrade to 3500 DC v3.1 to run data from the GlobalFiler Kits?
A: No, GlobalFiler/GlobalFiler Express Kit data can be run using Data Collection v1 but Data Collection v2 and Data Collection v3.1 offer multiple benefits to forensic laboratories including, but not limited to, Windows™ 7 compatibility, hard stop removals, increased polymer and buffer limits, 96-sample pouch support.
Q: If I upgrade to 3500 Data Collection v3.1 do I also need to upgrade my GeneMapper ID-X Software?
A: No, human identification (HID) sample files generated by the 3500 Data Collection software v2 can be viewed and analyzed with GeneMapper ID-X Software v1.2 and later. However, recommended version for Genemapper ID-X software is v1.5 that supports CODIS CMF v3.2 (R13) specification.
Q: If I upgrade my 3130 series instrument to run 6-dye data, can it still analyze 4- and 5-dye data?
A: Yes. The upgrade does not affect compatibility with current STR systems.
Q: If I upgrade to 3130/3130xl/3730 Data Collection Software v4, do I also need to upgrade my GeneMapper ID/ID-X Software?
A: If you upgrade to Data Collection v4 but do not activate the 6-dye module, you can analyze data using your current software version. If you have enabled the 6-dye module and are running GlobalFiler Kit data, the recommended version for GeneMapper ID-X Software is v1.5.
Q: How are previous versions of Data Collection upgraded?
A: The Data Collection Software upgrade is either performed by a field service engineer or can be performed by user depending on the version of the upgrade.
Upgrade from DC v1.0 to v3.1 is performed by a Field Service Engineer.
This upgrade involves the following steps:
Upgrading from DC v2.0 or v3.0 to v3.1 can be performed by the user.
Q: What is the new licensing scheme for the new versions of Data Collection Software?
A: To help us provide better communication on software updates or fixes, we use FlexNet™ licensing to register your Data Collection Software license. This model utilizes the computer’s ID and a license key to generate a unique installation ID to register the computer and activate the software at regular intervals.
A field service engineer installs the initial license and walks you through the registration process for future license renewals. A wizard within the Data Collection software will record the computer ID and
License Key, this information is used to register the license key at our registration website (the instrument computer DOES NOT need access to the Internet). The license registration process can be completed by transferring information via a thumb/flash drive. After registration is complete, a license file is sent to the PC (requires an Internet connection) and to an email address of your choice. If the instrument does not have Internet access, the user saves the license and returns to the License Activation Dialog within the Data Collection software to complete the installation and registration process. The data collection software license will need to be renewed every year. To renew the license, you will be prompted by the Data Collection software via the renewal process, which essentially requires you to resubmit the computer MAC ID and a license key to the registration site.
Q: What features are available in the GeneMapper ID-X Software v1.5?
|Microsoft® Windows® 7, 64-bit compatible||Supports this popular IT upgrade|
|Automated key software operations||Help reduce time and effort needed to transfer data to laboratory information management systems (LIMS) through automated export of individual sample plot PDFs and ability to create and split projects.|
|Support for CODIS CMF v3.2 (R13) specification||Helps reduce manual labor and transcription errors between GeneMapper® ID-X and CODIS upload.|
|Enhanced contamination assessment functionality||Helps enable a robust reference check with less effort through the ability to import legacy reference profiles and export comparison results for further data analysis and reporting.|
More detailed information on this software is available on the GeneMapper ID-X Software v1.5 User Bulletin.
Q: Can I upgrade from GeneMapper ID to GeneMapper ID-X v1.5?
A: There is no upgrade pathway from GeneMapper ID to GeneMapper ID-X v1.5. To start running GeneMapper ID-X Software v1.5 you will need at least 1 ‘Full Install’ version. To minimize the implementation and validation burden, GeneMapper ID-X software utilizes the same trusted and validated features and functionality of GeneMapper ID with additional features that specifically support the human identification (HID) data analysis workflow and the latest computer requirements of many laboratories. For more information on GeneMapper ID-X software features.
Q: I am using an earlier version of GeneMapper ID-X Software – how do I upgrade to GeneMapper ID-X Software v1.5?
A: You will need to install an upgrade license for each instance of GeneMapper ID-X you wish to upgrade. For example, if you have 1 Full GeneMapper ID-X v1.1 Software and 3 Client GeneMapper ID-X v1.1 Software licenses, you will need to install 1 License for GeneMapper ID-X v1.5 Full Upgrade Software and 3 Licenses of GeneMapper ID-X v1.5 Client Upgrade Software to upgrade all existing installations. Laboratories should run their own internal studies to confirm the performance of the software and if necessary, validate the optimal analysis parameters for their laboratory’s protocols.
Q: If I were to upgrade to GeneMapper ID-X Software v1.5, can I still view and analyze my old data?
A: Yes, GeneMapper ID-X Software v1.5 can analyze .fsa and .hid files generated from all Applied Biosystems™ Genetic Analyzers, all AmpFlSTR and GlobalFiler PCR Amplification Kits, as well as projects created using GeneMapper ID Software and previous versions of GeneMapper ID-X Software.
If you have additional comments or questions, please contact the customer service team via email.
If you are a current customer and need technical support, please contact the global service team via email, or phone us at 1-888-821-4HID (4443). Select option #1 for immediate assistance.
For Research Forensic or Paternity Use Only. Not for use in diagnostic procedures. For licensing and limited use restrictions visit thermofisher.com/HIDlicensing