Isolation of YAC insert sequences by representational difference analysis.

M Schutte; L T da Costa; C A Moskaluk; E Rozenblum; X Guan; P J de Jong; M Bittner; P S Meltzer; J M Trent; S E Kern

doi:10.1093/nar/23.20.4127

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PubMed · 7479076

Isolation of YAC insert sequences by representational difference analysis.

M Schutte·L T da Costa·C A Moskaluk·E Rozenblum·X Guan·P J de Jong·M Bittner·P S Meltzer·J M Trent·S E Kern

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Abstract

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.

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Base Sequence Carcinoma Chromosomes, Artificial, Yeast Chromosomes, Human, Pair 13 Cloning, Molecular DNA Dinucleotide Repeats Humans In Situ Hybridization, Fluorescence Molecular Sequence Data Pancreatic Neoplasms Polymerase Chain Reaction Sequence Tagged Sites Translocation, Genetic Tumor Cells, Cultured Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.

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M Schutte, L T da Costa, C A Moskaluk, E Rozenblum, X Guan, P J de Jong, M Bittner, P S Meltzer, J M Trent, S E Kern. 1995-10-25. Isolation of YAC insert sequences by representational difference analysis.. https://doi.org/10.1093/nar%2F23.20.4127

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