DNA probes: applications of the principles of nucleic acid hybridization.
Nucleic acid hybridization with a labeled probe is the only practical way to detect a complementary target sequence in a complex nucleic acid mixture. The first section of this article covers quantitative aspects of nucleic acid hybridization thermodynamics and kinetics. The probes considered are oligonucleotides or polynucleotides, DNA or RNA, single- or double-stranded, and natural or modified, either in the nucleotide bases or in the backbone. The hybridization products are duplexes or triplexes formed with targets in solution or on solid supports. Additional topics include hybridization acceleration and reactions involving branch migration. The second section deals with synthesis or biosynthesis and detection of labeled probes, with a discussion of their sensitivity and specificity limits. Direct labeling is illustrated with radioactive probes. The discussion of indirect labels begins with biotinylated probes as prototypes. Reporter groups considered include radioactive, fluorescent, and chemiluminescent nucleotides, as well as enzymes with colorimetric, fluorescent, and luminescent substrates.
Wetmur
J G
JG
Department of Microbiology, Mount Sinai School of Medicine, New York, New York.
D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D015342:DNA Probes / Q000096:biosynthesis / Q000138:chemical synthesis / Q000737:chemistry*; D004277:DNA, Single-Stranded; D007700:Kinetics; D008956:Models, Chemical; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D009692:Nucleic Acid Heteroduplexes; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000737:chemistry; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification; D015347:RNA Probes / Q000737:chemistry; D013816:Thermodynamics
D015342:DNA Probes; D004277:DNA, Single-Stranded; D009692:Nucleic Acid Heteroduplexes; D009841:Oligonucleotides; D015347:RNA Probes; D012313:RNA; D004247:DNA