Development of DNA-arrayed column for sensitive and selective analysis of DNA.
Quantitative separation of target DNA molecules was performed by DNA-arrayed silica capillary column on the basis of base pairing interaction of nucleic acids. We prepared DNA-arrayed silica capillary column by conjugating 5'-amniohexyl oligonucleotide (probe) on inner surface of the capillary column pre-treated with 3-aminopropyltriethoxysilane (APS) and disuccinimidyl glutarate (DSG) cross-linker. Sufficient resolution was observed by controlling of target-DNAs concentration, salt gradients, and temperature gradients. Finally, we succeeded to separate two different DNA targets even with same melting temperature according to their own concentrations. These results would be useful for developing quantitative analysis of cDNA, which is related to mRNA levels in biological sample.
Devarayapalli
Kamakshaiah Charyulu
KC
Institute of Advanced Energy, Kyoto University, Gokasho, Uji 611-0011, Japan.
Pack
Seung Pil
SP
Kamisetty
Nagendra Kumar
NK
Nonogawa
Mitsuru
M
Kodaki
Tsutomu
T
Makino
Keisuke
K
D002846:Chromatography, Affinity / Q000379:methods*; D004247:DNA / Q000032:analysis / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D015345:Oligonucleotide Probes; D012822:Silicon Dioxide / Q000737:chemistry
D015345:Oligonucleotide Probes; D012822:Silicon Dioxide; D004247:DNA