Synthesis and characterization of N‐rich fluorescent bio‐dots as a reporter in the design of dual‐labeled FRET probe for TaqMan PCR: A feasibility study. (17th August 2022)

• Record Type: Journal Article
• Authors: Imani, Mahsa; Mohajeri, Nasrin; Rastegar, Mojgan; Zarghami, Nosratollah
• Is Part Of: Biotechnology and applied biochemistry. Volume 70:Number 2(2023)
• Page Start: 645
• Page End: 658
• Publication Date: 2022-08-17
• Subjects: bioconjugation -- bio‐dot -- FRET probe; Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Genetic Techniques -- Periodicals; Microbiological Techniques -- Periodicals; 660.6
• DOI: 10.1002/bab.2387
• Languages: English
• ISSNs: 0885-4513

Abstract

Abstract: DNA‐based analytical techniques have provided an advantageous sensing assay in the realm of biotechnology. Bio‐inspired fluorescent nanodots are a novel type of biological staining agents with excellent optical properties widely used for cellular imaging and diagnostics. In the present research, we successfully synthesized bio‐dots with excellent optical properties and high‐quantum yield from DNA sodium salt through the hydrothermal method. We conjugated the bio‐dots with 3' Eclipse Dark Quencher (Eclipse)‐labeled single‐strand oligodeoxyribonucleotide according to carbodiimide chemistry, to design a fluorescence resonance energy transfer (FRET) probe. The results confirmed the prosperous synthesis and surface functionalization of the bio‐dot. Analysis of size, zeta potential, and FTIR spectroscopy verified successful bioconjugation of the bio‐dots with probes. UV‐visibility analysis and fluorescence intensity profile of the bio‐dot and bio‐dot@probes represented a concentration‐dependent quenching of fluorescent signal of bio‐dot by Eclipse after probe conjugation. The results demonstrated that TaqMan PCR was not feasible using the designed bio‐dot@probes. Our results indicated that bio‐dot can be used as an efficient fluorescent tag in the design of fluorescently labeled oligonucleotides with high biocompatibility and optical features. Abstract :