Optimization of Diamond Nucleic Acid Dye for Quantitative PCR

Alicia Haines, Shanan Tobe, Adrian Linacre

    Research output: Contribution to journalArticlepeer-review

    7 Citations (Scopus)

    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. Cqvalues 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.

    Original languageEnglish
    Pages (from-to)183-189
    Number of pages7
    JournalBiotechniques
    Volume61
    Issue number4
    DOIs
    Publication statusPublished - 2016

    Fingerprint Dive into the research topics of 'Optimization of Diamond Nucleic Acid Dye for Quantitative PCR'. Together they form a unique fingerprint.

    Cite this