STR profiling of epithelial cells identified by X/Y-FISH labelling and laser microdissection using standard and elevated PCR conditions

Lynch, Laura; Gamblin, Amelia; Vintiner, Sue; Simons, Joanne L.

doi:10.1016/j.fsigen.2014.10.017

Cited by 14 publications

(8 citation statements)

References 20 publications

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“…[49] used LCM to collect 15 to 150 FISH labelled diploid cells for direct PCR, using 28 cycles and the Identifiler®; PCR Amplification Kit, and investigated the heterozygote balance. Although one-tube extraction and direct PCR (i.e.…”

Section: Resultsmentioning

confidence: 99%

Characterization of degradation and heterozygote balance by simulation of the forensic DNA analysis process

2016

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Simulation experiments were used to show the impact of varying extraction efficiency, aliquot proportion, and PCR efficiency on the heterozygote balance of a range of diploid and haploid cells. Reducing either parameters introduces variance. It is well-known that the variance in heterozygote balance increases as the amount of DNA is reduced. Surprisingly the distribution is in fact diamond shaped — the variance start to decrease at very low amounts of DNA. Simulations suggest that pristine diluted DNA is an acceptable approximation in validations to infer heterozygote balance. However, the difference in distribution of the variance between diploid and haploid cell types may, under some circumstances, need to be considered in statistical models. Finally, we exemplify how simulations can be used to predict the outcome of PCR for degraded samples. Visualizing the predicted DNA profile as an electropherogram can help to identify the best approach for sample processing.Electronic supplementary materialThe online version of this article (doi:10.1007/s00414-016-1453-x) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Characterization of degradation and heterozygote balance by simulation of the forensic DNA analysis process

2016

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“…112 XY fluorescent in situ hybridization (FISH) coupled with laser microdissection (LMD) and DNA profiling at standard and low copy number conditions has been shown to be an effective way to obtain DNA profiling information from individual cell groups identified using specific fluorescent labeling techniques, in this case the X and Y chromosomes. 113 Detection of ESR1 (estrogen receptor 1) using fluorescently labeled monoclonal antibodies showed that buccal and vaginal epithelial cells were able to be distinguished using this marker when mRNA profiling could not. This shows that mRNA expression and protein expression in tissues do not always correlate.…”

Section: Dna-compatible Cell-specific Identificationmentioning

confidence: 99%

Forensic body fluid identification: state of the art

Harbison¹,

Fleming²

2016

RRFMS

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Body fluid identification is a key component in the forensic scientists' tool box and has been carried out both at the crime scene and in the laboratory for many years. Historically, methods relied on (bio) chemical-based tests, many of which lacked specificity. In this review, current technologies for identifying body fluids are described including the use of RNA (mRNA and miRNA), epigenetics, spectroscopic techniques such as Raman spectroscopy and microspectrophotometry, biosensors, and immunochromatographic methods which are outlined alongside their strengths and weaknesses. The potential for new insights into the identification of cells from new technologies such as massively parallel sequencing is explored.

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“…The FISH staining technique incorporates different colored DNA probes for particular parts of chromosomes in the nucleus. When coupled with laser microdissection, FISH can be an effective way to obtain DNA profiling information from specific cell groups that have been fluorescently labeled, such as those male cells containing X and Y chromosomes (Lynch, Gamblin, Vintiner, & Simons, ).…”

Section: Forensic Tests For Semenmentioning

confidence: 99%

The characteristics of seminal fluid and the forensic tests available to identify it

Gamblin

Morgan-Smith

2019

WIREs Forensic Science

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Semen body fluid identification remains a critical aspect of the forensic investigation of alleged sexual assaults. Over the years, as advancements have been made in the area of forensic science, several different methods for semen identification have been utilized. It is not unusual for several methods of semen identification to be incorporated into a single sexual assault investigation. Most methods of semen identification are based on either detecting enzymatic activities or are targeted at specific cells or antigens. Other methods have evolved utilizing fluorescence or protein expression. Presumptive tests are often used to screen or locate semen staining. Further confirmatory tests can be used to identify the staining as semen and finally the donor of the stain can be further identified through DNA analysis. For routine application of these methods, they need to be specific, sensitive, cost effective, easy to use, and nondestructive. Although new techniques are being researched, it appears that traditional methods of semen identification remain the most widely applied in forensic casework. This article is categorized under: Forensic Biology > Body Fluid Identification

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STR profiling of epithelial cells identified by X/Y-FISH labelling and laser microdissection using standard and elevated PCR conditions

Cited by 14 publications

References 20 publications

Characterization of degradation and heterozygote balance by simulation of the forensic DNA analysis process

Characterization of degradation and heterozygote balance by simulation of the forensic DNA analysis process

Forensic body fluid identification: state of the art

The characteristics of seminal fluid and the forensic tests available to identify it

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