Abstract
The GlobalFiler™ Express PCR Amplification Kit uses 6-dye fluorescent chemistry to
enable multiplexing of 21 autosomal STRs, 1 Y-STR, 1 Y-indel and the sex-determining
marker amelogenin. The kit is specifically designed for processing reference DNA samples
in a high throughput manner. Validation studies were conducted to assess the performance
and define the limitations of this direct amplification kit for typing blood and buccal
reference DNA samples on various punchable collection media. Studies included thermal
cycling sensitivity, reproducibility, precision, sensitivity of detection, minimum
detection threshold, system contamination, stochastic threshold and concordance. Results
showed that optimal amplification and injection parameters for a 1.2 mm punch from blood and buccal samples were 27 and 28 cycles, respectively, combined
with a 12 s injection on an ABI 3500xL Genetic Analyzer. Minimum detection thresholds were set
at 100 and 120 RFUs for 27 and 28 cycles, respectively, and it was suggested that data from positive
amplification controls provided a better threshold representation. Stochastic thresholds
were set at 250 and 400 RFUs for 27 and 28 cycles, respectively, as stochastic effects increased with cycle
number. The minimum amount of input DNA resulting in a full profile was 0.5 ng, however, the optimum range determined was 2.5–10 ng. Profile quality from the GlobalFiler™ Express Kit and the previously validated
AmpFlSTR® Identifiler® Direct Kit was comparable. The validation data support that
reliable DNA typing results from reference DNA samples can be obtained using the GlobalFiler™
Express PCR Amplification Kit.
Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Forensic Science International: GeneticsAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Genetics and genomics of core short tandem repeat loci used in human identity testing.J. Forensic Sci. 2006; 51: 253-265
- Development and validation of the AmpFlSTR Identifiler Direct PCR Amplification Kit: a multiplex assay for the direct amplification of single-source samples.J. Forensic Sci. 2011; 56: 835-845
- Direct STR amplification from whole blood and blood- or saliva-spotted FTA® without DNA purification.J. Forensic Sci. 2008; 53: 335-341
- Direct amplification of STRs from blood or buccal cell samples.Forensic Sci. Int. Genet. 2009; 2: 113-114
- Evaluation and comparative analysis of direct amplification of STRs using PowerPlex® 18D and Identifiler® direct systems.Forensic Sci. Int. Genet. 2012; 6: 640-645
- Expanding the CODIS core loci in the United States.Forensic Sci. Int. Genet. 2012; 6: e52-e54
- The evolution of DNA databases – recommendations for new European STR loci.Forensic Sci. Int. 2006; 156: 242-244
- Quality assurance standards for DNA databasing laboratories.Forensic Sci. Commun. 2000; 2 (Available at: http://www.fbi.gov/about-us/lab/biometric-analysis/codis/qas_databaselabs.pdf (accessed 07.15.13))
- GlobalFiler™ Express PCR Amplification Kit User Guide.Life Technologies, Carlsbad, CA2012
- PHASTR (Peak Height Analysis of STRs) Worksheet.2013 (Available at: http://web.unthsc.edu/info/200210/molecular_and_medical_genetics/887/research_and_development_laboratory/3)
- Developmental validation of the AmpFlSTR® NGM SElect™ PCR Amplification Kit: a next-generation STR multiplex with the SE33 locus.Forensic Sci. Int. Genet. 2013; 7: 41-51
- Run-specific limits of detection and quantitation for STR-based DNA testing.J. Forensic Sci. 2007; 52: 97-101
- Analytical thresholds and sensitivity establishing RFU thresholds for forensic DNA analysis.J. Forensic Sci. 2013; 58: 120-129
- Report for analytical chemists II. Quantitation in elemental analysis.Anal. Chem. 1970; 42: 26A-59A
- The low-template-DNA (stochastic) threshold – its determination relative to risk analysis for national DNA databases.Forensic Sci. Int. Genet. 2009; 3: 104-111
- Preservation and collection of biological evidence.Croat. Med. J. 2001; 42: 225-228
- Evaluation of saliva as a source of human DNA for population and association studies.Anal. Biochem. 2006; 353: 272-277
- Internal Validation Guide of Autosomal STR Systems for Forensic Laboratories.2013 (Available at: https://www.promega.com/∼/media/files/resources/validation%20guides/internal%20validation%20of%20autosomal%20str%20systems.pdf?la=en)
Article info
Publication history
Published online: January 31, 2014
Accepted:
January 18,
2014
Received in revised form:
January 14,
2014
Received:
December 5,
2013
Identification
Copyright
© 2014 Elsevier Ireland Ltd. Published by Elsevier Inc. All rights reserved.
