dna purification and analysis. the southern blot technique: -technique for dna analysis developed in...
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DNA Purification and Analysis
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The Southern Blot Technique:
-technique for DNA analysis developed in the 1970s by E. M. Southern
-Southern Blot allows detection and determination of the number of copies of particular sequences in complex DNA mixtures
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The detection of DNAs containing specific base sequencesby Southern Blotting
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-Restriction endonucleases are capableof selectively dissecting DNAmolecules into smaller fragments
-Restriction endonucleases permitconstruction of a new type of geneticmap, a restriction map, in which thesite of enzyme cleavage within theDNA is identified
-A comparison of restriction nucleasessites in mitochondrial DNA (mammalian mitochondrial DNA is acircular DNA molecule of 16,569 bp)allows for the calculation of the degreeof divergence in nucleotide sequencebetween species
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The Sanger Procedure of DNA Sequencing
-is based on the random termination of a DNA chain during enzymatic DNA synthesis
-the technique is possible because the dideoxynucleotide analog of each of the four normal nucleotides can be
incorporated into a growing DNA chain by DNA polymerase
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