Globalfiler

Manufactured by Thermo Fisher Scientific

Sourced in United States

The GlobalFiler is a DNA profiling kit designed for human identification analysis. It is a comprehensive solution for forensic and paternity testing applications. The kit enables the simultaneous amplification of multiple genetic markers, providing a high level of discrimination power for human identification.

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GlobalFiler® PCR Amplification Kit (21 citations) GlobalFiler™ Kit (3 citations) GlobalFiler STR Kit (2 citations) GlobalFiler™ IQC (2 citations) GlobalFiler Express PCR Amplification kit (2 citations)

6 protocols using globalfiler

Automated DNA Profiling from Blood Spots

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DNAs were punched and amplified from Nucleic-Cards (Copan, Italy) blood-spot samples using a fully automated workstation, starting from 1.2-mm diameter punches produced using the easyPunch STARlet system (Hamilton, Switzerland).
The samples were directly amplified using GlobalFiler (Thermo Fisher Scientific, Inc., Waltham, MA, USA) according to manufacturer’s recommendation. 15μl of low TE Buffer (pH 8.0) was added to the MicroAmp Optical 96-Well Reaction Plate (Thermo Fisher Scientific, Inc., Waltham, MA, USA) prior to the addition of 10μl of GlobalFiler master mix. A total of 24 loci were amplified, including 21 autosomal STR loci and three gender determination loci.
The PCR products (1μl) were separated by capillary electrophoresis in an ABI 3500xl Genetic Analyzer (Thermo Fisher Scientific Company, Carlsbad, USA) with reference to the LIZ600 size standard v2 (Thermo Fisher Scientific, Inc., Waltham, MA, USA) in total of 9 μl of LIZ600 standard and Hi-Di formamide (Thermo Fisher Scientific, Inc., Waltham, MA, USA) master mix. GeneMapper ID-X Software v1.4 (Thermo Fisher Scientific, Inc., Waltham, MA, USA) was used for genotype assignment. DNA typing and assignment of nomenclature were based on the ISFG recommendations.

Al-Snan N.R., Messaoudi S., R. Babu S, & Bakhiet M. (2019). Population genetic data of the 21 autosomal STRs included in GlobalFiler kit of a population sample from the Kingdom of Bahrain. PLoS ONE, 14(8), e0220620.

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Multiplex STR Genotyping for Human ID

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DNA samples were amplified with commercially available kits for human identification-oriented STRs markers Globalfiler (Thermo Fisher Scientific Inc.), PowerPlex Fusion (Promega Corporation, Madison, WI, United States) and PowerPlex Y23 (Promega Corporation), according to the manufacturer's instructions. [20] [21] [22] An additional analysis of chromosome 21 was performed by multiplex PCR, with a set of fluorescently labeled primers for the simultaneous amplification of four 21-STR loci: D21S1435 (21q21.1), D21S1437 (21q21.1), D21S2052 (21q21.1) and D21S2055 (21q22.2). Primer sequences are available in Supplementary Table 1. PCR was performed on Veriti 96-well Thermal Cycler equipment (Thermo Fischer Scientific Inc.) under following conditions: 95

Cavalheiro C.P., Avila E., Gastaldo A.Z., Graebin P., Motta C.H.A., Rodenbusch R, & Alho C.S. (2020). Uniparental disomy of chromosome 21: A statistical approach and application in paternity tests. Forensic science international. Genetics, 49.

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DNA Fragment Analysis via Genetic Analyzer

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One microliter of GlobalFiler™ or GlobalFiler™ Express (ThermoFisher Scientific, Carlsbad, CA, USA) amplified product was added to a mixture comprised of 9.5 μL Hi-Di™ formamide (ThermoFisher Scientific, Carlsbad, CA, USA) and 0.5 μL GeneScan™ 600 LIZ^® size standard (ThermoFisher Scientific, Carlsbad, CA, USA). Samples were injected on an Applied Biosystems’ 3500 Genetic Analyzer using POP-4™ polymer and Module J6 (15 s injection, 1.2 kV, 60 °C) and analyzed using GeneMapper™ ID-X v1.6 software (ThermoFisher Scientific, Carlsbad, CA, USA).

Huffman K., Hanson E, & Ballantyne J. (2022). Cell Subsampling Recovers Probative DNA Profile Information from Unresolvable/Undetectable Minor Donors in Mixtures. Genes, 13(7), 1117.

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PROVEDIt Dataset for Forensic STR Analysis

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The PROVEDIt dataset [7] 7.
Alfonse, L.E. IdentifilerPlus. The datasets were clustered into two sets (i.e., batches), with respect to those references that were contributors. Batch #1 consisted of two datasets, one with IdentifilerPlus (29 cycles) (Applied Biosystems) and the other with PowerPlex16 (Promega). Batch #2 consisted of three datasets, each with GlobalFiler (Thermo Fisher Scientific), Fusion 6C (Promega) and IdentifilerPlus (28 cycles), respectively. Different Applied Biosystems (ABI) CE instruments were used for the different datasets: ABI 3130 was used for the datasets with IdentifilerPlus (28 cycles) and PowerPlex16, whereas ABI 3500 was used for the others. Samples were run with several injection times, but for our study, the recommended injection times for each instrument were used: 5 s for 3130 and 15 s for 3500. The unfiltered version of the data (where all stutters are kept) was used (https://lftdi.camden.rutgers.edu/repository/PROVEDIt_1-5-Person%20CSVs%20UnFiltered.zip). All off-ladder (OL) alleles were removed. An overview of fragment length of each marker for the different multiplexes is provided in Fig. S1 (supplementary).

Bleka Ø., Prieto L, & Gill P. (2022). EFMrep: An extension of EuroForMix for improved combination of STR DNA mixture profiles. Forensic science international. Genetics, 61.

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GlobalFiler Amplification Protocol

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Amplification of the donor reference samples and the bulk DNA mixtures was conducted using the GlobalFiler^TM (ThermoFisher Scientific, Carlsbad, CA, USA) amplification kit according to the manufacturer’s recommended protocols. One nanogram of input DNA was used with an amplification protocol of 95 °C → 1 min; 29 cycles: 94 °C → 10 sec, 59 °C → 90 sec; 60 °C → 10 min; 4 °C → hold. Positive and negative amplification controls were included in each amplification.

Huffman K., Hanson E, & Ballantyne J. (2022). Cell Subsampling Recovers Probative DNA Profile Information from Unresolvable/Undetectable Minor Donors in Mixtures. Genes, 13(7), 1117.

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GlobalFiler DNA Amplification Protocol

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The GlobalFiler^TM (ThermoFisher Scientific, Carlsbad, CA, USA) amplification kit was used to amplify DNA from reference and bulk mixtures samples. One nanogram of input DNA was targeted, and the amplification protocol used was: 95 °C → 1 min; 29 cycles: 94 °C → 10 sec, 59 °C → 90 sec; 60 °C → 10 min; 4 °C → hold. Each amplification contained a positive and negative amplification control.

Huffman K, & Ballantyne J. (2022). Probabilistic Genotyping of Single Cell Replicates from Mixtures Involving First-Degree Relatives Prevents the False Inclusions of Non-Donor Relatives. Genes, 13(9), 1658.

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