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Second, because the bond cleaved by a restriction endonuclease is typically not positioned in the center of the recognition sequence to which it binds, and because the DNA strands are antiparallel, the cut sites for the two strands of a duplex are offset from each other (figure 2). After cleavage, each DNA fragment has a single-stranded end a few nucleotides long. The single-stranded ends of the two fragments are complementary to each other. Why Restriction Endonucleases Are So Useful There are hundreds of bacterial restriction endonucleases, and each one has a specific recognition sequence. By chance, a particular endonuclease's recognition sequence is likely to occur somewhere in any given sample of DNA; the shorter the sequence, the more often it will arise by chance within a sample.
Genetic engineering involves manipulating specific genes by cutting and rearranging DNA. A restriction endonuclease cleaves DNA at a specific site, generating in most cases two fragments with short single-stranded ends. Because these ends are complementary to each other, any pair of fragments produced by the same endonuclease, from any DNA source, can be joined together.
Many restriction endonucleases produce DNA fragments with "sticky ends." The restriction endonuclease EcoRI always cleaves the sequence GAATTC between G and A. Because the same sequence occurs on both strands, both are cut. However, the two sequences run in opposite directions on the two strands. As a result, single-stranded tails are produced that are complementary to each other, or "sticky."
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