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Artificial Cells, Nanomedicine, and Biotechnology
An International Journal
Volume 44, 2016 - Issue 3
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ORIGINAL ARTICLE

Sequence-specific label-free nucleic acid biosensor for the detection of the hepatitis C virus genotype 1a using a disposable pencil graphite electrode

Soner DonmezDepartment of Chemistry, Faculty of Science and Art, Nevsehir Haci Bektas Veli University, Nevsehir, TurkeyCorrespondence[email protected]
,
Fatma ArslanDepartment of Chemistry, Faculty of Science, Gazi University, Ankara, Turkey
&
Halit ArslanDepartment of Chemistry, Faculty of Science, Gazi University, Ankara, Turkey
Pages 912-917 | Received 20 Nov 2014, Accepted 11 Dec 2014, Published online: 24 Jan 2015

Figures & data

Table I. Oligonucleotide sequences employed (mismatches are indicated by bold and underlined characters).

Figure 1. Schematic diagram of experimental procedures.
Figure 1. Schematic diagram of experimental procedures.
Figure 2. Effect of probe concentration on the guanine oxidation signal: (a) after hybridization of complementary target oligonucleotide (cHCV1a), and (b) after hybridization of noncomplementary oligonucleotide (ncHCV3a). Error bars represent the standard deviation of three independent measurements.
Figure 2. Effect of probe concentration on the guanine oxidation signal: (a) after hybridization of complementary target oligonucleotide (cHCV1a), and (b) after hybridization of noncomplementary oligonucleotide (ncHCV3a). Error bars represent the standard deviation of three independent measurements.
Figure 3. Effect of the immobilization time on the guanine oxidation signal of the biosensor after hybridization with its complementary target oligonucleotide (c-HCV1a) and noncomplementary oligonucleotide (nc-HCV3a). Probe and target concentrations were 2.5 μM. Error bars represent the standard deviation of three independent measurements.
Figure 3. Effect of the immobilization time on the guanine oxidation signal of the biosensor after hybridization with its complementary target oligonucleotide (c-HCV1a) and noncomplementary oligonucleotide (nc-HCV3a). Probe and target concentrations were 2.5 μM. Error bars represent the standard deviation of three independent measurements.
Figure 4. Effect of the hybridization time on the guanine oxidation signal of the biosensor after hybridization with its complementary target oligonucleotide (c-HCV1a) and noncomplementary oligonucleotide (nc-HCV3a). Probe and target concentrations were 2.5 μM. Immobilization time was 60 min. Error bars represent the standard deviation of three independent measurements.
Figure 4. Effect of the hybridization time on the guanine oxidation signal of the biosensor after hybridization with its complementary target oligonucleotide (c-HCV1a) and noncomplementary oligonucleotide (nc-HCV3a). Probe and target concentrations were 2.5 μM. Immobilization time was 60 min. Error bars represent the standard deviation of three independent measurements.
Figure 5. (a) Effect of buffer solution on the hybridization reaction. (b) Effect of ionic strength on hybridization.
Figure 5. (a) Effect of buffer solution on the hybridization reaction. (b) Effect of ionic strength on hybridization.
Figure 6. Guanine oxidation signals versus hybridization temperature. Error bars represent the standard deviation of three independent measurements at each target DNA.
Figure 6. Guanine oxidation signals versus hybridization temperature. Error bars represent the standard deviation of three independent measurements at each target DNA.
Figure 7. Square wave voltammograms of guanine oxidation on the probe-immobilized PGE. (a) Complementary HCV1a (cHCV1a), noncomplementary HCV3a (ncHCV3a) and 3-base mismatch of HCV1a (3mm-HCV1a) oligonucleotide discrimination (b) Complementary HCV1a (RS-HCV1a) and noncomplementary HCV3a (RS-HCV3a) synthetic single-stranded PCR product analog discrimination.
Figure 7. Square wave voltammograms of guanine oxidation on the probe-immobilized PGE. (a) Complementary HCV1a (cHCV1a), noncomplementary HCV3a (ncHCV3a) and 3-base mismatch of HCV1a (3mm-HCV1a) oligonucleotide discrimination (b) Complementary HCV1a (RS-HCV1a) and noncomplementary HCV3a (RS-HCV3a) synthetic single-stranded PCR product analog discrimination.
Figure 8. Plot of the peak current vs. the concentration of HCV1a from 0.05 μM to 5 μM. Inset: related calibration graph at a concentration range of 0.05–0.75 μM.
Figure 8. Plot of the peak current vs. the concentration of HCV1a from 0.05 μM to 5 μM. Inset: related calibration graph at a concentration range of 0.05–0.75 μM.

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