GlobalFiler STR Kits
A 6-dye Chemistry offering superior discriminatory power
Around the world, forensic labs are asked to do more with less. That’s why Applied Biosystems GlobalFiler STR kits are tailored to combine maximum data recovery power with reduced amplification time. As part of the only fully integrated and validated forensic workflow, this 6-dye, 24-locus technology is designed to deliver superior lab performance—all facilitated by high-quality manufacturing at our ISO18385 certified facility, along with comprehensive training, service, and support.
Both the GlobalFiler and GlobalFiler Express kits are approved for use by laboratories generating DNA profiles for inclusion in the US National DNA Index System (NDIS) CODIS database.
Dr. Antonio Alonso discusses the Applied Biosystems 6-dye GlobalFiler kit, which has 5 orders of magnitude more discriminating power than previous kits and is very good for differentiating DNA mixtures.
Featured blog post and study: 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.
Featured technical documents
“Having 50% of the PCR products with a size less than or equal to 220 bp allows better amplification of degraded DNA. In addition, with a volume of 15 μ L of DNA per PCR reaction, we are in the best conditions for amplification of degraded DNA with an amount of DNA lower than100 pg/ μ L.”
“Seems to withstand well to inhibitors…”
“The baseline is perfect; no artifact was observed in negative controls. Sample profiles have showed good quality data with a good balance Peak Height Ratio (PHR).”
“Using a 6th dye allows to separate the markers analyzed and thus avoid allele overlap between neighboring markers.”
“The use of this kit should reduce the sample rate that has to be re-amplified whether with the same PCR kit or with another kit, while improving performance…”
“More sensitivity, more resistance to inhibition produces quality results.”
Need help bringing products online?
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.
- National Institute of Toxicology & Forensic Sciences, Madrid: Globalfiler Kit Validation
- Saint Louis County Police Crime Lab: GlobalFiler Kit Validation
- Saint Louis County Police Crime Lab: Interesting Cases
- Kansas City Police Crime Laboratory: Validation of GlobalFiler, Quantifiler on the 3500 GA
- HID Professional Services (HPS) tips and tricks for validations of the GlobalFiler™ Kit
- Upgrading the Workflow: Enhancing Information Recovery from Forensic Evidence
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.
For those situations where you seek increased sensitivity (such as when dealing with aged bone samples), the cycle number can be increased and a technical note is also 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
- Polymer and buffer limits* increased from 7 to 14 days, (8-capillary instrument) increased from 120 to 240 injections, (24-capillary instrument) increased from 50 to 100 injections
- 96 sample pouch support
- RFID limit updates (expiration date hard stop →warning changes)
- Microsoft™ Windows™ 7 Professional SP1 compatibility, 32 bit
- New PC (Dell™ Optiplex XE - 4 GB RAM, 500 GB RAID Disk)
- Modern Licensing Technology with a new license scheme that utilizes on-line registration
- Minor software updates
* 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:
- Set up new, Dell Optiplex XE Computer and Install OS Image
- Run a script to copy data from original computer
- Install the new Data Collection software, import data from original computer
- Run functional tests using Negative Controls to verify Data Collection is communicating properly with the instrument
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. Learn more ›
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. Learn more ›
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