Isolation of YAC insert sequences by representational difference analysis.
We present a method for the isolation of YAC insert sequences by representational difference analysis (RDA). To achieve maximal representation of the sequences, the amplicons were generated from a Mbol digestion product. RDA was performed using a 970 kb insert YAC clone. After two rounds of re-association and selective amplification 92% of the difference product represented sequences derived from the YAC insert. Twenty insert-specific sequence-tagged sites were readily defined. The difference product was also successfully used to isolate microsatellite markers, to identify clones from a human PAC library and as a chromosome painting probe in fluorescence in situ hybridization.
D001483:Base Sequence; D002277:Carcinoma; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D002882:Chromosomes, Human, Pair 13; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics*; D018900:Dinucleotide Repeats / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data; D010190:Pancreatic Neoplasms; D016133:Polymerase Chain Reaction / Q000379:methods; D016324:Sequence Tagged Sites; D014178:Translocation, Genetic / Q000235:genetics; D014407:Tumor Cells, Cultured
D004247:DNA