Massively parallel sequencing and capillary electrophoresis of a novel panel of falcon STRs: Concordance with minisatellite DNA profiles from historical wildlife crime
| Autor: | Guy Shorrock, Jordan Beasley, Celia A. May, Jon H. Wetton, Rita Neumann |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Massively parallel sequencing Wildlife laundering Raptor persecution Single-nucleotide polymorphism Locus (genetics) Peregrine Animals Wild Minisatellite Repeats Biology Polymorphism Single Nucleotide Pathology and Forensic Medicine DNA forensics 03 medical and health sciences 0302 clinical medicine Captive breeding Genetics Animals 030216 legal & forensic medicine Allele Massive parallel sequencing Electrophoresis Capillary High-Throughput Nucleotide Sequencing DNA Sequence Analysis DNA DNA Fingerprinting 030104 developmental biology Minisatellite DNA profiling Evolutionary biology Microsatellite Crime Research Paper STR multiplex Microsatellite Repeats |
| Zdroj: | Forensic Science International. Genetics |
| ISSN: | 1878-0326 1872-4973 |
| Popis: | Birds of prey have suffered persecution for centuries through trapping, shooting, poisoning and theft from the wild to meet the demand from egg collectors and falconers; they were also amongst the earliest beneficiaries of DNA testing in wildlife forensics. Here we report the identification and characterisation of 14 novel tetramer, pentamer and hexamer short tandem repeat (STR) markers which can be typed either by capillary electrophoresis or massively parallel sequencing (MPS) and apply them to historical casework samples involving 49 peregrine falcons, 30 of which were claimed to be the captively bred offspring of nine pairs. The birds were initially tested in 1994 with a multilocus DNA fingerprinting probe, a sex test and eight single-locus minisatellite probes (SLPs) demonstrating that 23 birds were unrelated to the claimed parents. The multilocus and SLP approaches were highly discriminating but extremely time consuming and required microgram quantities of high molecular weight DNA and the use of radioisotopes. The STR markers displayed between 2 and 21 alleles per locus (mean = 7.6), lengths between 140 and 360 bp, and heterozygosities from 0.4 to 0.93. They produced wholly concordant conclusions with similar discrimination power but in a fraction of the time using a hundred-fold less DNA and with standard forensic equipment. Furthermore, eleven of these STRs were amplified in a single reaction and typed using MPS on the Illumina MiSeq platform revealing eight additional alleles (three with variant repeat structures and five solely due to flanking SNPs) across four loci. This approach gave a random match probability of Graphical Abstract ga1 Highlights • Mini- and microsatellite DNA profiles identify false falcon captive breeding claims. • 12 novel tetra- to hexamer STRs used in capillary electrophoresis multiplexes. • First application of massively parallel sequencing to a wildlife crime case. • Massively parallel sequencing enhances discrimination by STR motif and SNP variants. • Impact of DNA testing on raptor wildlife crime and conservation. |
| Databáze: | OpenAIRE |
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