Response of DNA fragments to potentiometric sensors studied using HPLC.
Potentiometric sensors are studied as viable candidates for the construction of high throughput DNA arrays. For preliminary investigations, such sensors were used in an HPLC setup in the present work. This avoided errors due to ionic contaminants or additives in the commercial samples. The oligonucleotides dT(10), dT(20) and dT(30) were used as test substances. The potentiometric sensors were of the coated wire type, containing PVC, DOP, MTDDACl and a synthetic podand urea receptor. The HPLC system consisted of a reversed phase column eluted with a phosphate buffer, triethylammoniumacetate (TEAA), and an acetonitrile gradient. Molar responses and sensitivities increased with increasing chain length of oligonucleotides, yielding detection limits as low as 10(-6)M (dT(30), injected concentration). The slopes of the calibration graphs were at least 23 mV/decade (dT(10)), which was much higher than expected. The results are discussed in view of the potential use of this sensor type in high throughput microarrays.
Nagels
Luc J
LJ
Antwerp University, Chemistry Department, Groenenborgerlaan 171, B-2020, Antwerp, Belgium. luc.nagels@ua.ac.be
Everaert
Joseph
J
Bohets
Hugo
H
Del Favero
Jurgen
J
Goossens
Dirk
D
Robbens
Johan
J
Pietraszkiewicz
Marek
M
Pietraszkiewicz
Oksana
O
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D004566:Electrodes; D015394:Molecular Structure; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D009841:Oligonucleotides / Q000737:chemistry*; D011199:Potentiometry / Q000295:instrumentation / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D009841:Oligonucleotides