Direct PCR amplification of forensic touch and other challenging DNA samples: A review

Cavanaugh, Sarah E.; Bathrick, Abigail S.

doi:10.1016/j.fsigen.2017.10.005

Cited by 131 publications

(92 citation statements)

References 80 publications

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“…This time could be reduced further by utilising thermal cyclers with faster ramp rates, such as NextGen PCR technology (Molecular Biology Systems B. V.) [18]. Together with the increased use of direct PCR, not just for reference profiles but also touch and trace DNA [6,19], existing commercial forensic pipelines could be utilised to process both reference and casework samples in a rapid timescale using accredited facilities. In this study, we demonstrated the processing of FTA buccal cell samples in under 80 minutes, a time comparable to existing rapid DNA systems, but utilising existing commercial forensic laboratory pipelines with minor modifications to protocols.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to improvements made by STR kit manufacturers, published studies looking at reducing the time taken to generate a DNA profile using standard laboratory equipment have focused on use of direct PCR for touch and trace DNA [5,6], use of enhanced DNA polymerase enzymes [5,7,8,9,10], modified PCR mixes [9,11], and use of more efficient thermal cycler instrumentation [10,12]. In addition, research has also been carried out in the field of microfluidic chip systems to separate and detect fragments in place of traditional capillary electrophoresis platforms [5,13].…”

Section: Introductionmentioning

confidence: 99%

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Optimisation of rapid STR analysis using a standard DNA forensic pipeline

Gammon

Mayers

2019

Preprint

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Previous studies in published literature have reported on various alterations to STR mastermixes, protocols and instrumentation in order to reduce the time taken to generate forensic DNA profiles from reference and casework type samples. In this study, we demonstrate how altering default PCR amplification and capillary electrophoresis protocols in our existing DNA profiling pipeline can reduce the overall time taken to generate a DNA profile from buccal cell reference samples. GlobalFiler Express STR mastermix was used with direct PCR from FTA cards, run on altered PCR protocols and CE settings, and results compared to the standard evaluated settings used in our laboratories. This study demonstrated that full DNA profiles could be recovered in less than 80 minutes in comparison to our standard time of 97 -102 minutes whilst utilising existing reagent kits and instrumentation, with only minor modifications to protocols.Rapid DNA | direct amplification | forensic DNA analysis | STR profiling

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optimisation of rapid STR analysis using a standard DNA forensic pipeline

Gammon

Mayers

2019

Preprint

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“…The application of direct PCR in DNA profiling in forensic science has been reviewed by Cavanaugh and Bathrick (2018). Direct PCR has many advantages over the traditional approach.…”

Section: Dna Profilingmentioning

confidence: 99%

A review of direct polymerase chain reaction of DNA and RNA for forensic purposes

Lynch

Fleming

2019

WIREs Forensic Science

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Short tandem repeat (STR) profiling is frequently carried out on biological stains in forensic casework, and the application of messenger RNA (mRNA) body fluid identification is becoming more prevalent. Increasing constraints on analysis time, resources, an increase in the number of samples for analysis, and a large backlog of cases creates significant demand for quicker methods that direct polymerase chain reaction (PCR) can help overcome. Direct PCR bypasses DNA and/or RNA extraction, and in some cases, quantification by adding the sample or substrate punch to the amplification reaction. This method is currently used in some laboratories on reference samples. Direct PCR is also a component of portable devices, which could improve efficiency even further by producing results at the scene, and enabling triaging of samples for further testing. Portable devices are currently used for STR profiling of reference samples but not for mRNA profiling. Various concerns regarding the reliability of the method and equipment (portable devices) must first be addressed before incorporation into forensic casework. This review discusses the limitations and factors that need to be considered for direct PCR to be used for the analysis of casework samples, including being used for mRNA body fluid identification. This article is categorized under: Forensic Biology > Forensic DNA Technologies Forensic Biology > Body Fluid Identification Forensic Biology > Applications of RNA

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“…The uses of fluorophores sensitized by fluorescence resonance energy transfer (FRET) provided detection kinetics of fluorescence accumulation, and combined PCR amplification with real-time detection [76,77,99,100]. Alternative, or engineered, high-processivity DNA polymerases can have increased resistance to inhibitory factors relating to amplification of target DNA directly from samples without prior extraction or quantification [73,78,101,102].…”

Section: Extraction Of Compound And/or Challenging Samplesmentioning

confidence: 99%

Forensic DNA Technological Advancements as an Emerging Perspective on Medico-Legal Autopsy: A Mini Review

Pádár¹,

Zenke²,

Kozma³

2018

Post Mortem Examination and Autopsy - Current Issues From Death to Laboratory Analysis

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The importance of biological traces and evidences related to a criminal matter has been recognized for a long time. The examination of the expression of genetic polymorphism has been an integral part of the multidisciplinary field of medico-legal autopsy for over a century. Since the initial application of blood group antigens for personalization of a putative perpetrator in a murder case, the discipline of forensic genetics has evolved as a standard of forensic sciences. The real breakthrough, the application of molecular tools and processes for the in-vitro replication of genetic substances, has increasingly allowed the exploitation of advances of molecular genetics for both forensic and criminal investigations. Although there are certainly many more applications and scientific fields in the medico-legal arena, the relatively fast progress of genetics, which has accelerated recently with state-of-art technologies, can provide ever more relevant information in relation to a corpse or the cause and manner that resulted in the corpse for autopsy. This topic concerns the currently accepted forensic DNA technology, and the last section reviews commonly used markers for nuclear and mitochondrial DNA analysis as well as ongoing research. This review also focuses on the increasingly important non-human sources of DNA, and shortly covers the main aspects of animal forensic DNA examination.

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Direct PCR amplification of forensic touch and other challenging DNA samples: A review

Cited by 131 publications

References 80 publications

Optimisation of rapid STR analysis using a standard DNA forensic pipeline

Optimisation of rapid STR analysis using a standard DNA forensic pipeline

A review of direct polymerase chain reaction of DNA and RNA for forensic purposes

Forensic DNA Technological Advancements as an Emerging Perspective on Medico-Legal Autopsy: A Mini Review

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