DNA Purification and Analysis

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

The detection of DNAs containing specific base sequencesby Southern Blotting

-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

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