ORCID Profile
0000-0002-7380-2771
Current Organisation
Western Sydney University
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Publisher: Elsevier BV
Date: 12-2015
Publisher: Wiley
Date: 23-02-2015
Abstract: Nucleic acid staining dyes are used for detecting nucleic acids in electrophoresis gels. Historically, the most common dye used for gel staining is ethidium bromide, however due to its toxicity and mutagenicity other dyes that are safer to the user and the environment are preferred. This Short Communication details the properties of dyes now available and their sensitivity for detection of DNA and their ability to permeate the cell membrane. It was found that GelRed™ was the most sensitive and safest dye to use with UV light excitation, and both GelGreen™ and Diamond™ Nucleic Acid Dye were sensitive and the safer dyes using blue light excitation.
Publisher: Informa UK Limited
Date: 16-02-2019
Publisher: Future Science Ltd
Date: 10-2016
DOI: 10.2144/000114458
Abstract: Here, we evaluate Diamond Nucleic Acid Dye (DD) for use in quantitative PCR (qPCR) applications. Although DD is a commercially available stain for detection of DNA separated by gel electrophoresis, its use as a detection dye in qPCR has yet to be described. To determine if DD can be used in qPCR, we investigated its inhibitory effects on qPCR at concentrations ranging 0.1–2.5×. Serial dilution of DNA was used to determine the efficiency, sensitivity, and linearity of DD-generated qPCR data in comparison to other commonly used fluorescent dyes such as SYBR Green (SG), EvaGreen (EG), and BRYT Green (BG). DD was found to be comparable with other dyes for qPCR applications, with an R2 value .9 and an efficiency of 0.83. Mitochondrial DNA (mtDNA) target signals were successfully produced by DD over a DNA dilution range of ∼28 ng— 0.28 pg, demonstrating comparable sensitivity to the other dyes investigated. C q values obtained using DD were lower than those using EG by almost 7 cycles. We conclude that Diamond Nucleic Acid Dye is a cheaper, less toxic alternative for qPCR applications.
Publisher: Elsevier BV
Date: 05-2016
DOI: 10.1016/J.FORSCIINT.2016.03.026
Abstract: We report a simple screening method to assess the viability of successful DNA profiling from single hair follicles. A total of 48 hair s les (shed and plucked) were collected from male and female donors and the root tips (0.5cm) were stained using one of three DNA binding dyes (EvaGreen™, Diamond™ Nucleic Acid Dye and RedSafe™) at 20× concentration. The hairs were subsequently viewed under a Nikon Optiphot fluorescent microscope to count the approximate number of nuclei in one plane of view. The hairs were then processed using either (1) a DNA extraction kit (QIAmp(®) Mini Kit) and then lified using the AmpFLSTR(®) NGM™ kit, which lifies 15 short tandem repeat (STR) loci plus the gender marker amelogenin, or (2) by direct PCR lification using the same DNA profiling kit. Diamond™ dye had the lowest background signal and plucked hairs treated with this dye produced full DNA profiles when lified directly and was chosen to screen a further 150 mixed hair s les. These hairs were separated into one of five categories (1, >100 nuclei 1.5, 50-99 nuclei 2, 1-49 nuclei 2.5, no nuclei but high fluorescent signal 3, no nuclei and very low fluorescent signal) from which 60 of the hairs were chosen to undergo direct lification using the NGM™ kit. It was found that there was a direct correlation to the category designation and the ability to obtain a DNA profile up-loadable to the Australian DNA Database. Approximately 91% of category 1 hairs resulted in either a full or high partial (12-29 alleles) profile by direct PCR whereas about 78% of category 3 hairs exhibited no lification. The results show that this method can be used to predict successful STR lification from single hair follicles. It is a rapid, sensitive, cheap, non-destructive and easy to perform methodology applicable for screening multiple hairs in order to aid forensic investigators in predicting hairs that will yield DNA results.
Publisher: Elsevier BV
Date: 12-2015
Publisher: Informa UK Limited
Date: 02-01-2019
Publisher: Wiley
Date: 10-2015
Abstract: We report on the effects of six dyes used in the detection of DNA on the process of DNA extraction, lification, and detection of STR loci. While dyes can be used to detect the presence of DNA, their use is restricted if they adversely affect subsequent DNA typing processes. Diamond™ Nucleic Acid Dye, GelGreen™, GelRed™, RedSafe™, SYBR(®) Green I, and EvaGreen™ were evaluated in this study. The percentage of dye removed during the extraction process was determined to be: 70.3% for SYBR(®) Green I 99.6% for RedSafe™ 99.4% for EvaGreen™ 52.7% for Diamond™ Dye 50.6% for GelRed™, and could not be determined for GelGreen™. It was then assumed that the amount of dye in the fluorescent quantification assay had no effect on the DNA signal. The presence of all six dyes was then reviewed for their effect on DNA extraction. The t-test showed no significant difference between the dyes and the control. These extracts were then STR profiled and all dyes and control produced full DNA profiles. STR loci in the presence of GelGreen(TM) at 1X concentration showed increased lification products in comparison to the control s les. Full STR profiles were detected in the presence of EvaGreen™ (1X), although with reduced lification products. RedSafe™ (1X), Diamond™ Dye (1X), and SYBR(®) Green I (1X) all exhibited varying degrees of locus drop-out with GelRed™ generating no loci at all. We provide recommendations for the best dye to visualize the presence of DNA profile as a biological stain and its subsequent lification and detection.
Publisher: Elsevier BV
Date: 12-2015
Publisher: Elsevier BV
Date: 2013
Location: New Zealand
No related grants have been discovered for Alicia Haines.