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Updated: Jul 5, 2026
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In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
Published on: March 25, 2020
Design, synthesis, and amplification of DNA pools for in vitro selection
J Pollard1, S D Bell, A D Ellington
1Harvard University, Cambridge, Massachusetts, USA.
Current Protocols in Nucleic Acid Chemistry
|April 23, 2008
Abstract:
Preparation of a random-sequence DNA pool is presented. The degree of randomization and the length of the random sequence are discussed, as is synthesis of the pool using a DNA synthesizer. Purification of a single-stranded pool and conversion to a double-stranded pool are presented as step-by-step protocols. Support protocols describe determination of the complexity and skewing of the pool, and optimization of amplification conditions.
