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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.

No MeSH data available.


Frequency response of QCM electrode modified with probe 1 via avidin-biotin system in the presence of:(a)complementary PCR product - fragment of EPSPS gene (2.5 nM)(b)non-complementary PCR product - fragment of Adh gene (2.5 nM)The solution composition: 27 mM HEPES, 55 mM NaCl, 0.05 mM EDTA, 2.5 mM MgCl2, pH 7.7; total volume in the QCM cell: 200 μl.
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f5-sensors-07-01462: Frequency response of QCM electrode modified with probe 1 via avidin-biotin system in the presence of:(a)complementary PCR product - fragment of EPSPS gene (2.5 nM)(b)non-complementary PCR product - fragment of Adh gene (2.5 nM)The solution composition: 27 mM HEPES, 55 mM NaCl, 0.05 mM EDTA, 2.5 mM MgCl2, pH 7.7; total volume in the QCM cell: 200 μl.

Mentions: As a next step the sensor was used for the detection of PCR amplified 169 base pairs fragment of EPSPS gene. The electrode modified with probe 1 was more sensitive and in the presence of 2.5 nM EPSPS-derived oligonucleotide a -88.2 ± 7.5 Hz frequency change was observed (Figure 5, curve a).


Piezoelectric Sensor for Determination of Genetically Modified Soybean Roundup Ready ® in Samples not Amplified by PCR
Frequency response of QCM electrode modified with probe 1 via avidin-biotin system in the presence of:(a)complementary PCR product - fragment of EPSPS gene (2.5 nM)(b)non-complementary PCR product - fragment of Adh gene (2.5 nM)The solution composition: 27 mM HEPES, 55 mM NaCl, 0.05 mM EDTA, 2.5 mM MgCl2, pH 7.7; total volume in the QCM cell: 200 μl.
© Copyright Policy
Related In: Results  -  Collection

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

f5-sensors-07-01462: Frequency response of QCM electrode modified with probe 1 via avidin-biotin system in the presence of:(a)complementary PCR product - fragment of EPSPS gene (2.5 nM)(b)non-complementary PCR product - fragment of Adh gene (2.5 nM)The solution composition: 27 mM HEPES, 55 mM NaCl, 0.05 mM EDTA, 2.5 mM MgCl2, pH 7.7; total volume in the QCM cell: 200 μl.
Mentions: As a next step the sensor was used for the detection of PCR amplified 169 base pairs fragment of EPSPS gene. The electrode modified with probe 1 was more sensitive and in the presence of 2.5 nM EPSPS-derived oligonucleotide a -88.2 ± 7.5 Hz frequency change was observed (Figure 5, curve a).

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.