Revision of the nonequilibrium thermal dissociation and stringent washing approaches for identification of mixed nucleic acid targets by microarrays.

摘要: Microarray experiments typically involve washing steps that remove hybridized nonspecific targets with the purpose of improving signal-to-noise ratio. The quality ultimately affects downstream analysis microarray and interpretation. paucity fundamental studies directed towards understanding dissociation mixed from microarrays makes development meaningful washing/dissociation protocols difficult. To fill void, we examined activation energies preexponential coefficients 47 perfect match (PM) double-mismatch (MM) duplex pairs to discover there was no statistical difference between kinetics PM MM duplexes. Based on these findings, evaluated nonequilibrium thermal (NTD) approach, which has been used identify specific microbial in target samples. We found major premises for various NTD approach might be seriously compromised because: (i) duplexes do not always dissociate before ones, (ii) relationship rates depends temperature sequence. Specifically NTD, show previously suggested use reference curves, indices curves ramps lead erroneous conclusions.