Journal article
Optimization of diamond nucleic acid dye for quantitative PCR
BioTechniques, Vol.61(4)
2016
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 >0.9 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. Cq 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.
Details
- Title
- Optimization of diamond nucleic acid dye for quantitative PCR
- Authors/Creators
- A.M. Haines (Author/Creator)S.S. Tobe (Author/Creator)A. Linacre (Author/Creator)
- Publication Details
- BioTechniques, Vol.61(4)
- Publisher
- Informa BioSciences
- Identifiers
- 991005541656007891
- Copyright
- © 2016 Future Science Group
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- International collaboration
- Citation topics
- 1 Clinical & Life Sciences
- 1.54 Molecular & Cell Biology - Genetics
- 1.54.1454 PCR Techniques
- Web Of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology
- ESI research areas
- Biology & Biochemistry