Temperature effects on DNA chip experiments from surface plasmon resonance imaging: isotherms and melting curves.
We present an analysis of hybridization experiments on a DNA chip studied by surface plasmon resonance imaging. The reaction constants at various temperatures and for different probe lengths are obtained from Langmuir isotherms and hybridization kinetics. The melting curves from temperature scans are also obtained without any labeling of the targets. The effects of the probe length on the hybridization thermodynamics, deduced from the temperature dependence of the reaction constants as well as from the melting curves, suggest dispersion in the length of the hybridization segments of the probes accessible to the targets. Those are, however, sufficient to suggest efficient point mutation detection from temperature scans.
Fiche
J B
JB
SPrAM UMR 5819, Département de Recherche Fondamentale sur la Matière Condensée, CEA Grenoble, France.
Buhot
A
A
Calemczuk
R
R
Livache
T
T
D016477:Artifacts; D004247:DNA / Q000032:analysis / Q000737:chemistry* / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017403:In Situ Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D044367:Phase Transition; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance / Q000379:methods*; D013696:Temperature; D044366:Transition Temperature
D004247:DNA