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An approach to DNA sequencing in which genetic markers are mapped and DNA sequences are aligned by matching overlapping sites of known sequence. |
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Any of a type of enzyme that cleaves double-stranded DNA at specific sites; extensively used in recombinant DNA technology. Also called a restriction endonuclease. |
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A relatively rapid method of DNA sequencing in which a DNA molecule is broken up into overlapping fragments, each fragment sequenced, and then analyzed |
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Single Nucleotide Polymorphism |
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Inherited variation in a single nucleotide base in DNA that differ between individuals. |
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Bacterial artificial chromosome (BAC) |
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A DNA cloning vector used in bacteria that can carry up to 150,000 base pairs of foreign DNA. |
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All of the cloned DNA fragments generated by the action of a restriction endonuclease on a genome. |
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A relatively rapid method of DNA sequencing in which a DNA molecule is broken up into overlapping fragments, each fragment is sequenced, and high-speed computers analyze and realign the fragments |
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High-throughput sequencing |
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Rapid DNA sequencing on a micro scale in which many fragments of DNA are sequenced in parallel |
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To determine the sequence of a DNA fragment it is isolated and mixed with |
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Definition
DNA polymerase A short primer appropriate for the DNA sequence The four dNTPs (dATP, dGTP, dCTP, and dTTP) Small amounts of the four ddNTPs, each bonded to a differently colored fluorescent “tag”
deoxyribonucleoside triphosphates (dNTPs) |
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Figure 17.1 Sequencing Genomes Involves Fragment overlaps |
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Shows how each of the fragments have to be aligned according to the areas in which they overlap. Both Hierarchical sequencing and shotgun sequencing are used. |
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Figure 17.2 Sequencing DNA |
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The normal substrates for DNA replication are dNTPs. The chemically modified structure of ddNTPs causes DNA synthesis to stop. When labeled ddNTPs are incorporated into a reaction mixture for replicating a DNA template of unknown sequence, the result is a collection of fragments of varying lengths that can be separated by electrophoresis. |
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Figure 17.3 High-Throughput Sequencing |
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High-speed sequencing is faster and cheaper than traditional methods, and involves the chemical amplification of DNA fragments. In this illustration Fluorescent base is added and analyzed by a computer. |
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Figure 17.4 The Genomic book of life |
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Genome sequences contain many features, some of which are summarized in this overview. Sifting through all the information contained in a genome sequence can help us understand how an organism functions and what its evolutionary history might be. |
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