Padlock probes: circularizing oligonucleotides for localized DNA detection.
Nucleotide sequence information derived from DNA segments of the human and other genomes is accumulating rapidly. However, it frequently proves difficult to use such short DNA segments to identify clones in genomic libraries or fragments in blots of the whole genome or for in situ analysis of chromosomes. Oligonucleotide probes, consisting of two target-complementary segments, connected by a linker sequence, were designed. Upon recognition of the specific nucleic acid molecule the ends of the probes were joined through the action of a ligase, creating circular DNA molecules catenated to the target sequence. These probes thus provide highly specific detection with minimal background.
Nilsson
M
M
Beijer Laboratory, Department of Medical Genetics, Biomedical Center, Uppsala, Sweden.
Malmgren
H
H
Samiotaki
M
M
Kwiatkowski
M
M
Chowdhary
B P
BP
Landegren
U
U
D001483:Base Sequence; D002478:Cells, Cultured; D002881:Chromosomes, Human, Pair 12; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA / Q000032:analysis*; D004270:DNA, Circular / Q000032:analysis*; D005822:Genetic Vectors; D006801:Humans; D017403:In Situ Hybridization; D008214:Lymphocytes; D008565:Membrane Proteins / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes / Q000737:chemistry; D012091:Repetitive Sequences, Nucleic Acid; D013698:Templates, Genetic
C505032:CFTR protein, human; D004270:DNA, Circular; D008565:Membrane Proteins; D015345:Oligonucleotide Probes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA