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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.


TEM image of (A) water soluble CdSe QD and (B) CdSe QD-ssDNA conjugate with magnification of 200,000×.
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biosensors-03-00419-f003: TEM image of (A) water soluble CdSe QD and (B) CdSe QD-ssDNA conjugate with magnification of 200,000×.

Mentions: The particle size and morphology of water soluble CdSe QD and CdSe QD-ssDNA conjugate were analyzed using TEM. Figure 3 shows TEM image for both samples in 100 nm scale. It can be viewed that the size of the particles were in the range of 2 to 10 nm. These results also proved that the modification and conjugation of QD did not affect its particle size. It can been seen that in Figure 3(A), a single particle of QD is observed while in Figure 3(B), agglomeration is observed possibly due to conjugation with ssDNA.


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)

TEM image of (A) water soluble CdSe QD and (B) CdSe QD-ssDNA conjugate with magnification of 200,000×.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

biosensors-03-00419-f003: TEM image of (A) water soluble CdSe QD and (B) CdSe QD-ssDNA conjugate with magnification of 200,000×.
Mentions: The particle size and morphology of water soluble CdSe QD and CdSe QD-ssDNA conjugate were analyzed using TEM. Figure 3 shows TEM image for both samples in 100 nm scale. It can be viewed that the size of the particles were in the range of 2 to 10 nm. These results also proved that the modification and conjugation of QD did not affect its particle size. It can been seen that in Figure 3(A), a single particle of QD is observed while in Figure 3(B), agglomeration is observed possibly due to conjugation with ssDNA.

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.