EZ-Tn5™ Transposase completely sequences cDNA or genomic clones in plasmid, cosmid, fosmid, or BAC vectors without subcloning or primer walking.
EZ-Tn5™ Transposase is a hyperactive form of Tn5 transposase.1 The highly purified, single-subunit enzyme can be used to randomly insert (transpose or "hop") any EZ-Tn5 Transposon into any target DNA in vitro with an efficiency up to >106 insertion clones per standard reaction. When incubated with an EZ-Tn5 Transposon in the absence of Mg2 , a stable EZ-Tn5 Transposome™ complex is formed. The Transposome is so stable that it can be electroporated into living cells. Once in the cell, the Transposome is activated by intracellular Mg2 and the EZ-Tn5 Transposon component is randomly inserted into the host's genomic DNA. The three-dimensional structure of a Transposome complex has been elucidated.2
A typical EZ-Tn5 transposition reaction requires four components: (1) the EZ-Tn5 Transposase; (2) an EZ-Tn5 Transposon; (3) a target DNA; and (4) the presence of Mg2 . The highly random insertion of an EZ-Tn5 Transposon into the target DNA3 proceeds by a cut-and-paste mechanism,1 catalyzed by the EZ-Tn5 Transposase, and results in a 9-bp duplication of target DNA sequence immediately adjacent to both ends of the Transposon.
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A typical EZ-Tn5™ transposition reaction requires four components:
- EZ-Tn5™ Transposase
- EZ-Tn5™ Transposon
- a target DNA
Insertion of an EZ-Tn5™ Transposon into the target DNA proceeds by a cut and paste mechanism,1 catalysed by the EZ-Tn5™ Transposase, and results in a 9-bp duplication of target DNA sequence immediately adjacent to both ends of the Transposon.
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