High-throughput analysis using AmpFlSTR® Identifiler® with the Applied Biosystems 3500xl Genetic Analyser
| Autor: | Amanda Kirkham, Yordina Haile, C. P. Kimpton, Andrea Grout, Peter Gill, John Patrick Haley, Ahmed Al-Marzouqi |
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| Rok vydání: | 2012 |
| Předmět: |
Heterozygote
Analyser Bioinformatics Risk Assessment Pathology and Forensic Medicine 03 medical and health sciences 0302 clinical medicine Limit of Detection Genetics Humans Detection theory 030216 legal & forensic medicine Dropout (neural networks) 030304 developmental biology Mathematics Detection limit 0303 health sciences Stochastic Processes Homozygote Gene Amplification High-Throughput Nucleotide Sequencing DNA High throughput analysis DNA profiling Biological system Microsatellite Repeats |
| Zdroj: | Forensic science international. Genetics. 7(1) |
| ISSN: | 1878-0326 |
| Popis: | High throughput analysis of buccal scrape reference samples using the Identifiler system on the Applied Biosystems 3500 xl Genetic Analyser is described. This platform is much more sensitive than previous platforms, e.g. 3130 xl . The range of signal detection is also much greater and this means that the system is more tolerant to a wide range of input template concentrations. DNA quantity is not a limiting factor in the analysis of buccal scrapes, hence the entire analytical procedure (process) was designed around a target input of 1.5ng, to minimise low level DNA profiles and associated events of allele dropout. A universal stochastic threshold/limit of detection (LOD) of 300relative fluorescent units (rfu) was applied across all loci. This level is twice that of comparable systems such as SGM Plus analysed on 3130 xl instruments. After analysis of dropout probabilities, heterozygous balance, drop-in and stutter characteristics, rule-sets were programmed into the FSS i-cubed software. The process has a success rate >95%. No discordances or mis-designations occurred when applied to a validation set of more than 1000 samples. |
| Databáze: | OpenAIRE |
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