Amplified oligonucleotide sensing in microliter volumes containing copper ions by solution streaming.
We present a simple, cost-effective design for amplifying oligodeoxynucleotide (ODN) sensing, in microliter ODN volumes containing copper ions, by solution streaming (bubbling). The inert gas streaming (bubbling) at a constant pressure of 0.04 bar drives the motion of a 30-microL ODN droplet containing a three-electrode circuit (inverted drop microcell), and in the presence of copper ions offers an approximately 50-times improvement in the detection of ODN samples. The detection of ODNs at the carbon paste electrode is based on the enhancement of the oxidation peaks of purine bases (adenine and guanine) by the anodic stripping of the electrochemically accumulated complex of Cu(I) with purine base residues of acid hydrolyzed ODN samples (Cu(I)-ahODN complex). We used the proposed method for (i) the determination of the percentage content of adenine and guanine units within analyzed ODN samples at subnanomolar concentrations (related to monomer content) and (ii) the detection of the (TTC)n triplet expansion using magnetic DNA hybridization with reporter probes containing guanine units (the TTC trinucleotide repeat expansion is associated with serious hereditary diseases, including Friedreich ataxia).
Hason
Stanislav
S
Institute of Biophysics, Academy of Sciences of the Czech Republic, Kralovopolska 135, CZ-612 65 Brno, Czech Republic. hasons@ibp.cz
Vetterl
Vladimír
V
D000225:Adenine / Q000737:chemistry; D001128:Argon / Q000737:chemistry; D001483:Base Sequence; D003300:Copper / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics; D006147:Guanine / Q000737:chemistry; D007477:Ions / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D009841:Oligonucleotides / Q000737:chemistry* / Q000235:genetics; D012996:Solutions
D015342:DNA Probes; D007477:Ions; D009841:Oligonucleotides; D012996:Solutions; D006147:Guanine; D001128:Argon; D003300:Copper; D000225:Adenine