Fundamentals of DNA Separation and Detection

Butler, John M.

doi:10.1016/b978-0-12-374999-4.00009-6

Cited by 14 publications

(20 citation statements)

References 38 publications

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“…Unlike commercial capillary electrophoresis (CE), the development of CE on novel systems such as the one described herein inherently requires that a number of parameters be optimized for effective functioning of the system [ 44 ]. Previous work has shown sufficient separation of 10 STR loci ranging from 80 to 500 bp using this on-disc platform [ 30 , 36 ], thus, in this study we focused on optimizing sample input, heating of the separation channel, and sample injection time to adequately separate the amplicons in three body fluid panels.…”

Section: Resultsmentioning

confidence: 99%

Rapid Microchip Electrophoretic Separation of Novel Transcriptomic Body Fluid Markers for Forensic Fluid Profiling

et al. 2022

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Initial screening of criminal evidence often involves serological testing of stains of unknown composition and/or origin discovered at a crime scene to determine the tissue of origin. This testing is presumptive but critical for contextualizing the scene. Here, we describe a microfluidic approach for body fluid profiling via fluorescent electrophoretic separation of a published mRNA panel that provides unparalleled specificity and sensitivity. This centrifugal microfluidic approach expedites and automates the electrophoresis process by allowing for simple, rotationally driven flow and polymer loading through a 5 cm separation channel; with each disc containing three identical domains, multi-sample analysis is possible with a single disc and multi-sample detection per disc. The centrifugal platform enables a series of sequential unit operations (metering, mixing, aliquoting, heating, storage) to execute automated electrophoretic separation. Results show on-disc fluorescent detection and sizing of amplicons to perform comparably with a commercial ‘gold standard’ benchtop instrument and permitted sensitive, empirical discrimination between five distinct body fluids in less than 10 min. Notably, our microfluidic platform represents a faster, simpler method for separation of a transcriptomic panel to be used for forensically relevant body fluid identification.

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Section: Resultsmentioning

confidence: 99%

Rapid Microchip Electrophoretic Separation of Novel Transcriptomic Body Fluid Markers for Forensic Fluid Profiling

et al. 2022

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“…repeated in variable numbers among different individuals that can be informative for discrimination of identities [8], a significant aspect in forensic investigation. In addition, DNA profiling has also been utilized to resolve paternity and kinship issues, development of DNA database search, and forensic intelligence [1,9]. In this context, the utilization of autosomal Short Tandem Repeats (STRs) has acquired considerable attention due to its evidential value in establishing individualities [10].…”

Section: X-str Profiling and Its Evidential Valuesmentioning

confidence: 99%

“…While an autosomal STR refers to a marker on 'a chromosome not involved in sex determination', an X-chromosomal STR (X-STR) is 'the marker found on the X chromosome that can sometimes be useful in addressing kinship analysis' [9]. The utilization of X-STRs DNA profiling is deemed important in complementing the autosomal STRs DNA profiling, particularly in complex kinship cases (e.g.…”

Section: X-str Profiling and Its Evidential Valuesmentioning

confidence: 99%

Applications of X-Chromosome Short Tandem Repeats for Human Identification: A Review

Mahat

Alwi²,

Salleh³

et al. 2023

J.Trop.Life.Science

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The forensic DNA profiling technique has tremendously contributed to forensic human identification, an important aspect in forensic investigations. In instances whereby comparison samples are unavailable, utilization of short tandem repeats of X chromosome (X-STRs) may prove useful to resolve complex kinship investigations involving missing persons and mass disasters. Despite such evidential values, the use of X-STRs during investigations remains scarce in many Southeast Asian countries including Malaysia, requiring concerted efforts for establishing forensic statistical support for its diverse populations (especially the admixture populations), standardizing core loci and procedure, improving the knowledge among practitioners as well as developing suitable standard operating procedure for incorporating X-STRs analysis in the overall DNA profiling framework. Hence, this review paper aims to highlight the developments, applications and population data of X-STRs, as well as its challenges and future insights for forensic casework.

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“…However it involves the use of some potentially toxic substances and can contaminate proteins [24]. Using traditional methods, such as phenolchloroform, teeth samples can be challenging to process, resulting in low-quantity and/or quality nuclear DNA and insufficient profiles for comparisons [25]. Both methods were used to evaluate the DNA yield in a resource limited setting.…”

Section: Deoxyribonucleic Acid (Dna) Extractionmentioning

confidence: 99%

“…The low yield of DNA in both methods used in this study maybe connected to the presence co-extracting impurities from accelerants and the high calcium content due to the use of the whole tooth which can act as PCR inhibitors [28]. In the study of Edson [25], which used teeth samples tested in AmpFlSTR_ MiniFiler, a silica-based extraction kit, 16/70 teeth tested positive for STR loci, of which only 4 loci were positively identified.…”

Section: Deoxyribonucleic Acid (Dna) Extractionmentioning

confidence: 99%

The Integrity of Deoxyribonucleic Acid (DNA) Extracted from Whole Teeth Samples Burnt with Different Accelerants Using Two Extraction Protocols for Forensic Sex Determination

Iroanya¹,

Akinshipo²,

Mairiga³

2022

EQUIJOST

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Sex determination is usually the first step in forensic identification of victim(s) in disasters, jungle justice and arsonscenarios for many medical and legal reasons. The use of DNA in forensic analysis offers a good method in sexdetermination and the quality of extracted DNA is very important for downstream PCR. The aim of this study is toinvestigate the viability of DNA obtained from burnt teeth for forensic sex-determination. Two DNA extractionmethods consisting of silica based commercial kit and phenol-chloroform organic method, followed by polymerasechain amplification of amelogenin gene for sex determination were employed. Based on the DNA yield and opticaldensity values, the kit extraction method performed better than the phenol-chloroform method with 100% and85.71% success respectively. A Mann-Whitney U test of 260/280 absorbance showed no statistically significantdifference in the median absorbance for aviation fuel (median = 1.34) and gasoline (median =1.32) burnt samples.Amplification of AMEL genes using the commercial kit and phenol-chloroform method were 52.38% and 22.22%positive respectively compared to pre-extraction sex determination. This study shows that the use of the silica basedcommercial kit technique yielded higher DNA quality and quantity from whole teeth burnt with gasoline andaviation fuel as accelerants for downstream PCR amplification of AMEL genes compared to organic phenolchloroform method.

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Fundamentals of DNA Separation and Detection

Cited by 14 publications

References 38 publications

Rapid Microchip Electrophoretic Separation of Novel Transcriptomic Body Fluid Markers for Forensic Fluid Profiling

Rapid Microchip Electrophoretic Separation of Novel Transcriptomic Body Fluid Markers for Forensic Fluid Profiling

Applications of X-Chromosome Short Tandem Repeats for Human Identification: A Review

The Integrity of Deoxyribonucleic Acid (DNA) Extracted from Whole Teeth Samples Burnt with Different Accelerants Using Two Extraction Protocols for Forensic Sex Determination

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