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For example, a library of fragments of a coding sequence can be generated by: treating a double-stranded PCR fragment of the coding sequence of interest with a nuclease under conditions in which nicking occurs only about once per molecule, denaturing the double-stranded DNA, renaturing the DNA to form double-stranded DNA that may include sense/antisense pairs from different nicking products, removing single-stranded portions from the reformed duplex by treatment with S1 nuclease, and ligating the resulting fragment library to an expression vector

G., Potenza, F., Viele, M., Dufrusine, B., Franciotti, R., Pietrangelo, L., Ardini, M., Stuppia, L., Federici, L., De Laurenzi, V., & Sallese, M