AI Article Synopsis
- The study evaluates the effectiveness of Diamond Nucleic Acid Dye (DD) as a detection dye in quantitative PCR (qPCR), which hasn't been reported before.
- DD showed comparable performance to other fluorescent dyes like SYBR Green, EvaGreen, and BRYT Green in terms of efficiency, sensitivity, and linearity for qPCR applications.
- The findings suggest that DD is a cost-effective and less toxic alternative for detecting DNA in qPCR, with better sensitivity indicated by lower cycle threshold (Cq) values compared to EvaGreen.
Article 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.
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| http://dx.doi.org/10.2144/000114458 | DOI Listing |
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