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BIOL 3110
Peter Cheung

Oct 1, 2009 Restriction Mapping -This process of creating a restriction map is based on the idea that there are different recognition sites for the various other types of endonucleases. -Some of the restriction enzymes will leave “sticky” or “staggered” ends that allow for the creation of recombinant DNA molecules (the staggered ends of each piece allows for modified reannealing). -There are three types of restriction enzymes (I, II, III) but only type II is used for mapping. Type I does not create DNA fragments reproducibly; type III can cut reproducibly but has a cognate methylase associated with type I and forms a complex when in the same solution. -Type II is the only useful one in recombinant technology because it cuts reproducibly (restriction enzyme activity requires Mg and cleaves the phosphate bond) and because it does not have a methylase associated with the other types of restrictions enzymes; it is also the only one that creates staggered ends during the cutting process. 5’ Protruding Ends/Tails 4 5’ – G(3’OH) 4 (5’PO )AATC – 3’ 3’ – CTTAA(5’PO ) (3’OH)G – 5’ -the staggered ends are palindromic and allow for the reannealing of the ends, the conditions for reannealing are different from those of digestion. Palindrome Sequences & Mapping DNA Sequences -Differ in length and sequence of recognition sites; the shorter recognition sites (i.e. 4bp vs. 6bp)
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