Genetic profiling from challenging samples: Direct PCR of touch DNA

Templeton, Jennifer E.L.; Ottens, Renée; Paradiso, Viviana; Handt, Oliva; Taylor, Duncan; Linacre, Adrian

doi:10.1016/j.fsigss.2013.10.115

Cited by 37 publications

(22 citation statements)

References 3 publications

(3 reference statements)

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“…Samples and substrates used in this study did not reflect all possible scenarios of contact DNA left behind in forensic investigations (e.g., damaged, degraded, and intact cellular DNA). We have previously reported that the direct PCR method can be used to generate full DNA profiles from fingermarks deposited 2 h after individuals washed their hands (15). The direct PCR approach in this study led to significantly higher peak heights of alleles (p = 0.001) compared to the extraction experiment when controlling for DNA mass and substrate (See Table S1).…”

Section: Discussionmentioning

confidence: 72%

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Direct PCR Improves the Recovery of DNA from Various Substrates

Templeton

Taylor

Handt

et al. 2015

Journal of Forensic Sciences

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This study reports on the comparison of a standard extraction process with the direct PCR approach of processing low-level DNA swabs typical in forensic investigations. Varying concentrations of control DNA were deposited onto three commonly encountered substrates, brass, plastic, and glass, left to dry, and swabbed using premoistened DNA-free nylon FLOQswabs(™) . Swabs (n = 90) were either processed using the DNA IQ(™) kit or, for direct PCR, swab fibers (~2 mm(2) ) were added directly to the PCR with no prior extraction. A significant increase in the height of the alleles (p < 0.005) was observed when using the direct PCR approach over the extraction methodology when controlling for surface type and mass of DNA deposited. The findings indicate the potential use of direct PCR for increasing the PCR product obtained from low-template DNA samples in addition to minimizing contamination and saving resources.

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Section: Discussionmentioning

confidence: 72%

“…However, in light of recent advances, new swab types and new swab media can now be considered as alternative methods for DNA recovery. Novel DNA capture methods in conjunction with direct PCR can be used to explore the potential for improvement in information obtained from forensic samples .…”

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confidence: 99%

Direct PCR Improves the Recovery of DNA from Various Substrates

Templeton

Taylor

Handt

et al. 2015

Journal of Forensic Sciences

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“…Our method is rapid (23) because there is no need for a DNA extraction step, and there is a reduction in associated costs because there is no need to purchase DNA extraction kits. Most importantly for forensic science laboratories, by eliminating the need to increase PCR cycle number or concentrate the amplified products, the procedure is easily adapted into working practices, allowing laboratories accredited to ISO17025 to use the method.…”

Section: Resultsmentioning

confidence: 99%

“…If DNA exists as a free molecule, then it should bind readily to positively charged fibers (23). Furthermore, the use of a surfactant such as Triton-X is thought to assist in the binding of DNA to a swab (24) and hence, these two features were used to capture DNA from touched substrates.…”

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confidence: 99%

DNA Profiles from Fingermarks

Templeton

Linacre

2014

BioTechniques

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Criminal investigations would be considerably improved if DNA profiles could be routinely generated from single fingermarks. Here we report a direct DNA profiling method that was able to generate interpretable profiles from 71% of 170 fingermarks. The data are based on fingermarks from all 5 digits of 34 individuals. DNA was obtained from the fingermarks using a swab moistened with Triton-X, and the fibers were added directly to one of two commercial DNA profiling kits. All profiles were obtained without increasing the number of amplification cycles; therefore, our method is ideally suited for adoption by the forensic science community. We indicate the use of the technique in a criminal case in which a DNA profile was generated from a fingermark on tape that was wrapped around a drug seizure. Our direct DNA profiling approach is rapid and able to generate profiles from touched items when current forensic practices have little chance of success.

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“…Alternatively, swabs or other materials can be used to remove touch DNA from a substrate and then the swab head can be introduced directly into the PCR reaction. The successful use of direct amplification for touch DNA recovery has been widely reported from a range of sample types [7][8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Touch DNA on objects can be analysed at low cost using simplified direct amplification methods

Gammon

Murray-Jones

Shenton

et al. 2019

Preprint

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Previous studies reported in the literature demonstrate that a range of sampling vehicles can be used effectively for forensic analysis of human DNA in direct amplification reactions. In this study we compared Copan microFLOQ ® swabs with a range of alternative sampling vehicles, using touch DNA samples donated by 15 different volunteers. MicroFLOQ swabs performed well, as did 3 mm diameter discs punched from analytical filter paper. The 3 mm discs could be used in a 5 µl PCR volume, increasing sensitivity, and reducing costs when compared with other methods that require a larger PCR volume. Other inert sampling vehicles, such as interdental toothbrushes and toothpicks also gave good results in direct amplification. The study found a large variation in results between the 15 touch DNA donors, demonstrating the importance of validating touch DNA recovery techniques with a large pool of donors.touch DNA | direct amplification | forensic DNA analysis | trace evidence

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Genetic profiling from challenging samples: Direct PCR of touch DNA

Cited by 37 publications

References 3 publications

Direct PCR Improves the Recovery of DNA from Various Substrates

Direct PCR Improves the Recovery of DNA from Various Substrates

DNA Profiles from Fingermarks

Touch DNA on objects can be analysed at low cost using simplified direct amplification methods

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