Sensitive quantitative nucleic acid detection using oligonucleotide microarrays.
We report a new theoretical approach to optimize the performance and quantify the results of gene expression oligonucleotide microarrays which are widely used in biomedical research. An on-array hybridization isotherm that takes into account the screened Coulomb repulsion between the assayed nucleic acid target and the layer of surface tethered oligonucleotide probes is presented. The hybridization efficiency is found as a function of the genomic target (sequence, length, and concentration), array parameters (probe sequence and length, surface probe density), and hybridization conditions (temperature and buffer ionic strength). We present simple relations for the hybridization signal maximum and the linear dynamic detection range and show explicit criteria for optimization. The approach is based on an extension of our recently published theory (Vainrub, A.; Pettitt, B. M. Phys. Rev. E 2002, 66, art. no.-041905) which we generalize here for the cases of target depletion effects and arbitrary target length.
Vainrub
Arnold
A
Department of Chemistry, University of Houston, Houston, TX 77204-5003, USA.
Pettitt
B Montgomery
BM
D009696:Nucleic Acids / Q000032:analysis* / Q000737:chemistry; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis / Q000737:chemistry; D012680:Sensitivity and Specificity; D055672:Static Electricity; D013816:Thermodynamics
D009696:Nucleic Acids; D012333:RNA, Messenger