Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2824456 | Plasmid | 2008 | 8 Pages |
Abstract
We have developed and tested a method for the restriction enzyme-independent generation of sticky-end PCR products. The method is suitable for use with a proof-reading polymerase such as pfu, or any other heat-stable polymerase which produces a blunt-end product. The technique can be used to achieve unidirectional cloning of PCR products with an efficiency greater than 90%. Because the sequences of the sticky ends are defined by the user and potentially can be of any length, the method can also be exploited for the one-step construction of recombinant plasmids from multiple functional cassettes, without the use of restriction enzymes.
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Authors
Andrew Walker, James Taylor, Duncan Rowe, David Summers,