Constructing contigs from large-insert clones.
This unit describes three approaches that are widely used to define alignments between overlapping clones bearing large-insert genomic DNA and to generate extensive contiguous overlapping sets of clones (contigs). The three approaches are sequence-tagged site (STS) content mapping, repetitive-element hybridization fingerprinting, and Alu-PCR fingerprinting. Methods for isolating the necessary BAC DNA suitable for automated fluorescent sequencing and generating new STS markers are discussed in support protocols. An alternate protocol presents repetitive-element hybridization fingerprinting to detect overlaps and build contigs with full-genomic YAC libraries.
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Walter and Eliza Hall Institute, Victoria, Australia.
Marshall
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Munroe
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D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D020451:Contig Mapping / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D016172:DNA Fingerprinting; D005826:Genetics, Medical; D015698:Genomic Library; D006801:Humans; D016133:Polymerase Chain Reaction; D016324:Sequence Tagged Sites
D004247:DNA