Generating DNA profile from low copy number DNA: Strategies and associated risks

Mateen, Rana Muhammad; Tariq, Asma; Hussain, Mureed

doi:10.4025/actascibiolsci.v42i1.52239

Cited by 2 publications

(2 citation statements)

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“…STR artifacts such as allele drop-out, stutter product formation, allele drop-in, allele slippage, and other microvariations are outcomes of various types of genetic variations taking place at the molecular level in vivo [6,7]. Some of the STR profiles of earwax samples, after 60 days of storage at room temperature, showed locus drop-out, a single allele dropout or drop-in, or a stutter in others.…”

Section: Discussionmentioning

confidence: 99%

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Short Tandem Repeat (STR) Profiling of Earwax DNA Obtained from Healthy Volunteers

Amer,

Alotaibi,

Shahid

et al. 2023

CIMB

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The present study aimed to establish human earwax as a potential source of DNA evidence that could be effectively used in human identification. Sixty earwax samples were obtained from 15 healthy male and female Saudi volunteers living in Riyadh, Saudi Arabia. Four consecutive earwax swab samples were obtained from each volunteer and stored for 1, 15, 30 and 60 days. Earwax samples were stored at room temperature (20–22 °C). Reference oral swab was also taken from each volunteer. DNA was extracted by QIAamp DNA Mini kit and quantified by real-time polymerase chain reaction (RT-PCR) on 7500 Thermal Cycler. Autosomal STR loci were amplified using AmpFLSTR™ Identifiler™ Plus PCR Amplification Kit (Thermo Fisher Scientific, Carlsbad, CA, USA). Amplified fragments were size separated and analyzed on a 3500 Genetic Analyzer. Complete autosomal STR profiles were obtained from the earwax swabs of all the volunteers stored up to 30 days after the collection. Some STR profiles were partially obtained 60 days after the earwax collection. Allelic drop-out, allelic drop-in, and stutters were seen in earwax samples analyzed 60 days after the collection. The results have shown that human earwax can be a potential source of DNA evidence for human identification up to 30 days after the earwax collection. It is recommended to quickly analyze earwax samples or store them at room temperature or at −10 °C after their recovery from the crime scene.

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Section: Discussionmentioning

confidence: 99%

“…Of the various artifacts produced by degraded earwax DNA typing, allelic drop-out, allelic drop-in, locus drop-out, and stutters are quite common [6,7]. Allele drop-out is an extreme example of heterozygote imbalance, where one allele falls below the limit of the detection threshold (LDT) [8].…”

Section: Introductionmentioning

confidence: 99%