The use of oligodeoxynucleotide probes in chaotrope-based hybridization solutions.
Hybridization solutions containing chaotropes may be used to modulate the thermal stability (Tm or Td) of oligodeoxynucleotide (ODN) duplexes or hybrids over a 90 degrees C range. Modulation of Td allows formulation of hybridization solutions that permit ambient temperature hybridization using most combinations of probe length, probe composition, target type, and facilitates development of convenient and rapid assay formats. The conditions required to achieve ODN duplex fidelity, and optimal yields of hybridized product, are described for trichloroacetate, thiocyanate, guanidinium salts and other chaotropic salts. The effects of different solid supports on Td are described. Also, a method is presented that uses chaotropic compounds to reduce background arising from signal ODN probes in a sandwich assay hybridization format.
Van Ness
J
J
MicroProbe Corporation, Bothell, WA 98021.
Chen
L
L
D001483:Base Sequence; D006146:Guanidines / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D012996:Solutions / Q000737:chemistry*; D013696:Temperature; D013861:Thiocyanates / Q000378:metabolism; D014238:Trichloroacetic Acid / Q000378:metabolism
D006146:Guanidines; D015345:Oligonucleotide Probes; D012996:Solutions; D013861:Thiocyanates; D012965:Sodium Chloride; D014238:Trichloroacetic Acid; C031760:thiocyanate