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Piezoelectric Sensor for Determination of Genetically Modified Soybean Roundup Ready ® in Samples not Amplified by PCR

View Article: PubMed Central

ABSTRACT

The chemically modified piezoelectrodes were utilized to develop relatively cheap and easy to use biosensor for determination of genetically modified Roundup Ready soybean (RR soybean). The biosensor relies on the immobilization onto gold piezoelectrodes of the 21-mer single stranded oligonucleotide (probes) related to 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene, which is an active component of an insert integrated into RR soybean genome. The hybridization reaction between the probe and the target complementary sequence in solution was monitored. The system was optimized using synthetic oligonucleotides, which were applied for EPSPS gene detection in DNA samples extracted from animal feed containing 30% RR soybean amplified by the PCR and nonamplified by PCR. The detection limit for genomic DNA was in the range of 4.7·105 numbers of genom copies contained EPSPS gene in the QCM cell. The properties such as sensitivity and selectivity of piezoelectric senor presented here indicated that it could be applied for the direct determination of genetically modified RR soybean in the samples non-amplified by PCR.

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Cyclic voltammetry reductive desorption of 3,3′-dithiodipropionic acid di(N-succinimidyl ester) from gold piezoelectrode after two hours of modification in 5 mM of chloroform solution. Measurement conditions: 0.5 M KOH, potential scan rate 100 mV/s.This was sufficient for avidin immobilization. The presence of 0.2 mg ·ml-1 avidin in the QCM cell resulted in approximately 100 Hz decrease of piezoelectrode frequency. The immobilization of avidin on the electrode surface via creation of amide bonds with 3,3′-dithiodipropionic acid di-(N-succinimidyl ester) was completed after 30 min.(Figure 3).
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f2-sensors-07-01462: Cyclic voltammetry reductive desorption of 3,3′-dithiodipropionic acid di(N-succinimidyl ester) from gold piezoelectrode after two hours of modification in 5 mM of chloroform solution. Measurement conditions: 0.5 M KOH, potential scan rate 100 mV/s.This was sufficient for avidin immobilization. The presence of 0.2 mg ·ml-1 avidin in the QCM cell resulted in approximately 100 Hz decrease of piezoelectrode frequency. The immobilization of avidin on the electrode surface via creation of amide bonds with 3,3′-dithiodipropionic acid di-(N-succinimidyl ester) was completed after 30 min.(Figure 3).

Mentions: Figure 1 illustrates the multistep process of the modification of piezoelectrodes. The presence of 3,3′-dithiodipropionic acid di-(N-succinimidyl ester) on the electrode surface was confirmed by performing the reductive desorption process. It is known that the potential cycled from -0.4V to -1.2V in alkaline solution (0.5 M KOH) disrupts Au-S covalent bonds [43, 44]. The amount of adsorbed ester may be estimated from the charge required for reductive desorption. The density of 3,3′-dithiodipropionic acid di-(N-succinimidyl ester) on the electrode surface after 2h modification was calculated from the area of the reduction peak (Figure 2), and equaled - 18.8 × 1010 molecules·mm-2.


Piezoelectric Sensor for Determination of Genetically Modified Soybean Roundup Ready ® in Samples not Amplified by PCR
Cyclic voltammetry reductive desorption of 3,3′-dithiodipropionic acid di(N-succinimidyl ester) from gold piezoelectrode after two hours of modification in 5 mM of chloroform solution. Measurement conditions: 0.5 M KOH, potential scan rate 100 mV/s.This was sufficient for avidin immobilization. The presence of 0.2 mg ·ml-1 avidin in the QCM cell resulted in approximately 100 Hz decrease of piezoelectrode frequency. The immobilization of avidin on the electrode surface via creation of amide bonds with 3,3′-dithiodipropionic acid di-(N-succinimidyl ester) was completed after 30 min.(Figure 3).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3814863&req=5

f2-sensors-07-01462: Cyclic voltammetry reductive desorption of 3,3′-dithiodipropionic acid di(N-succinimidyl ester) from gold piezoelectrode after two hours of modification in 5 mM of chloroform solution. Measurement conditions: 0.5 M KOH, potential scan rate 100 mV/s.This was sufficient for avidin immobilization. The presence of 0.2 mg ·ml-1 avidin in the QCM cell resulted in approximately 100 Hz decrease of piezoelectrode frequency. The immobilization of avidin on the electrode surface via creation of amide bonds with 3,3′-dithiodipropionic acid di-(N-succinimidyl ester) was completed after 30 min.(Figure 3).
Mentions: Figure 1 illustrates the multistep process of the modification of piezoelectrodes. The presence of 3,3′-dithiodipropionic acid di-(N-succinimidyl ester) on the electrode surface was confirmed by performing the reductive desorption process. It is known that the potential cycled from -0.4V to -1.2V in alkaline solution (0.5 M KOH) disrupts Au-S covalent bonds [43, 44]. The amount of adsorbed ester may be estimated from the charge required for reductive desorption. The density of 3,3′-dithiodipropionic acid di-(N-succinimidyl ester) on the electrode surface after 2h modification was calculated from the area of the reduction peak (Figure 2), and equaled - 18.8 × 1010 molecules·mm-2.

View Article: PubMed Central

ABSTRACT

The chemically modified piezoelectrodes were utilized to develop relatively cheap and easy to use biosensor for determination of genetically modified Roundup Ready soybean (RR soybean). The biosensor relies on the immobilization onto gold piezoelectrodes of the 21-mer single stranded oligonucleotide (probes) related to 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene, which is an active component of an insert integrated into RR soybean genome. The hybridization reaction between the probe and the target complementary sequence in solution was monitored. The system was optimized using synthetic oligonucleotides, which were applied for EPSPS gene detection in DNA samples extracted from animal feed containing 30% RR soybean amplified by the PCR and nonamplified by PCR. The detection limit for genomic DNA was in the range of 4.7·105 numbers of genom copies contained EPSPS gene in the QCM cell. The properties such as sensitivity and selectivity of piezoelectric senor presented here indicated that it could be applied for the direct determination of genetically modified RR soybean in the samples non-amplified by PCR.

No MeSH data available.


Related in: MedlinePlus