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Development of a Fluorescence Resonance Energy Transfer (FRET)-Based DNA Biosensor for Detection of Synthetic Oligonucleotide of Ganoderma boninense.

Mohd Bakhori N, Yusof NA, Abdullah AH, Hussein MZ - Biosensors (Basel) (2013)

Bottom Line: Hybridization time, temperature and effect of different concentration of target DNA were studied in order to optimize the developed system.TEM results show that the particle size of QD varies in the range between 5 to 8 nm after ligand modification and conjugation with ssDNA.This approach is capable of providing a simple, rapid and sensitive method for detection of related synthetic DNA sequence of Ganoderma boninense.

View Article: PubMed Central - PubMed

Affiliation: Chemistry Department, Faculty Science, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia; E-Mails: noremyliamb@gmail.com (N.M.B.); halim@science.upm.edu.my (A.H.A.); mzobir@science.upm.edu.my (M.Z.H.).

ABSTRACT
An optical DNA biosensor based on fluorescence resonance energy transfer (FRET) utilizing synthesized quantum dot (QD) has been developed for the detection of specific-sequence of DNA for Ganoderma boninense, an oil palm pathogen. Modified QD that contained carboxylic groups was conjugated with a single-stranded DNA probe (ssDNA) via amide-linkage. Hybridization of the target DNA with conjugated QD-ssDNA and reporter probe labeled with Cy5 allows for the detection of related synthetic DNA sequence of Ganoderma boninense gene based on FRET signals. Detection of FRET emission before and after hybridization was confirmed through the capability of the system to produce FRET at 680 nm for hybridized sandwich with complementary target DNA. No FRET emission was observed for non-complementary system. Hybridization time, temperature and effect of different concentration of target DNA were studied in order to optimize the developed system. The developed biosensor has shown high sensitivity with detection limit of 3.55 × 10(-9) M. TEM results show that the particle size of QD varies in the range between 5 to 8 nm after ligand modification and conjugation with ssDNA. This approach is capable of providing a simple, rapid and sensitive method for detection of related synthetic DNA sequence of Ganoderma boninense.

No MeSH data available.


Fluorescence spectra of (a) CdSe quantum dot (QD)-ssDNA, (b) CdSe QD-ssDNA hybridized with complementary DNA and (c) CdSe QD-ssDNA hybridized with non-complementary DNA; Excitation was fixed at 488 nm; scan rate: 2 nm/s; slidwidth: 10 nm.
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biosensors-03-00419-f001: Fluorescence spectra of (a) CdSe quantum dot (QD)-ssDNA, (b) CdSe QD-ssDNA hybridized with complementary DNA and (c) CdSe QD-ssDNA hybridized with non-complementary DNA; Excitation was fixed at 488 nm; scan rate: 2 nm/s; slidwidth: 10 nm.

Mentions: The changes of the fluorescence signals for CdSe QD-ssDNA conjugate and hybrid system contained target DNAs (complementary and non-complementary DNA) were used to characterize the sensing capability of the developed detection system. Figure 1 shows fluorescence spectra of CdSe QD-ssDNA and CdSe QD-dsDNA hybridized with complementary and non-complementary target DNA. The emission peak at 640 nm, which corresponds to the emission of QD, was obtained for all spectra. Only hybridization with complementary target DNA shows emission peak at 680 nm confirming the occurrence of hybridization.


Development of a Fluorescence Resonance Energy Transfer (FRET)-Based DNA Biosensor for Detection of Synthetic Oligonucleotide of Ganoderma boninense.

Mohd Bakhori N, Yusof NA, Abdullah AH, Hussein MZ - Biosensors (Basel) (2013)

Fluorescence spectra of (a) CdSe quantum dot (QD)-ssDNA, (b) CdSe QD-ssDNA hybridized with complementary DNA and (c) CdSe QD-ssDNA hybridized with non-complementary DNA; Excitation was fixed at 488 nm; scan rate: 2 nm/s; slidwidth: 10 nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263569&req=5

biosensors-03-00419-f001: Fluorescence spectra of (a) CdSe quantum dot (QD)-ssDNA, (b) CdSe QD-ssDNA hybridized with complementary DNA and (c) CdSe QD-ssDNA hybridized with non-complementary DNA; Excitation was fixed at 488 nm; scan rate: 2 nm/s; slidwidth: 10 nm.
Mentions: The changes of the fluorescence signals for CdSe QD-ssDNA conjugate and hybrid system contained target DNAs (complementary and non-complementary DNA) were used to characterize the sensing capability of the developed detection system. Figure 1 shows fluorescence spectra of CdSe QD-ssDNA and CdSe QD-dsDNA hybridized with complementary and non-complementary target DNA. The emission peak at 640 nm, which corresponds to the emission of QD, was obtained for all spectra. Only hybridization with complementary target DNA shows emission peak at 680 nm confirming the occurrence of hybridization.

Bottom Line: Hybridization time, temperature and effect of different concentration of target DNA were studied in order to optimize the developed system.TEM results show that the particle size of QD varies in the range between 5 to 8 nm after ligand modification and conjugation with ssDNA.This approach is capable of providing a simple, rapid and sensitive method for detection of related synthetic DNA sequence of Ganoderma boninense.

View Article: PubMed Central - PubMed

Affiliation: Chemistry Department, Faculty Science, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia; E-Mails: noremyliamb@gmail.com (N.M.B.); halim@science.upm.edu.my (A.H.A.); mzobir@science.upm.edu.my (M.Z.H.).

ABSTRACT
An optical DNA biosensor based on fluorescence resonance energy transfer (FRET) utilizing synthesized quantum dot (QD) has been developed for the detection of specific-sequence of DNA for Ganoderma boninense, an oil palm pathogen. Modified QD that contained carboxylic groups was conjugated with a single-stranded DNA probe (ssDNA) via amide-linkage. Hybridization of the target DNA with conjugated QD-ssDNA and reporter probe labeled with Cy5 allows for the detection of related synthetic DNA sequence of Ganoderma boninense gene based on FRET signals. Detection of FRET emission before and after hybridization was confirmed through the capability of the system to produce FRET at 680 nm for hybridized sandwich with complementary target DNA. No FRET emission was observed for non-complementary system. Hybridization time, temperature and effect of different concentration of target DNA were studied in order to optimize the developed system. The developed biosensor has shown high sensitivity with detection limit of 3.55 × 10(-9) M. TEM results show that the particle size of QD varies in the range between 5 to 8 nm after ligand modification and conjugation with ssDNA. This approach is capable of providing a simple, rapid and sensitive method for detection of related synthetic DNA sequence of Ganoderma boninense.

No MeSH data available.