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 >10 6 insertion clones per standard reaction. When incubated with an EZ-Tn5 Transposon in the absence of Mg 2+, 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 Mg 2+ 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
Applications
In vitro insertion of an EZ-Tn5 Transposon into DNA cloned in vectors, such as plasmids, fosmids, cosmids, or BACs as well as in vitro insertion into linear DNA.*
Preparation of EZ-Tn5 Transposomes for in vivo transposition following electroporation into living cells.**
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 Mg 2+. The highly random insertion of an EZ-Tn5 Transposon into the target DNA 3 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.
Benefits
Specifically and uniquely recognises the Outer End sequences of naturally occurring Tn5 and mini-Tn5 transposons and the hyperactive Mosaic Ends of EZ-Tn5 Transposons.
References
Goryshin, I.Y. and Reznikoff , W.S. (1998) J. Biol. Chem. 273, 7367.
Davies, D.R. et al. (2000) Science 289, 77.
Shevchenko, Y. et al. (2002) Nucleic Acids Res. 30, 2469.
*Covered byissued and/or pending patents, exclusively licensed or assigned to Epicentre® (an Illumina® Company).
**The use of Transposome™ complexes for in vivo insertion of a transposon, including, but not limited, to HyperMu™ and EZ-Tn5™ Transposome™ complexes, is covered by U.S. Patent No. 6,159,736 and related patents and patent applications, exclusively licensed to Epicentre® (an Illumina® Company).
Key features
- Create custom transposomes for in vitro transposomics
- Integrate selectable markers, promoters or replication origins into DNA in vitro with EZ-Tn5 insertion kits
Product Details
10 units of EZ-Tn5 transposase is enough enzyme for 10 standard reactions.
Please note: Illumina requires that a Transposome License Acknowledgement is signed prior to processing orders for EZ-Tn5 Transposase. Please complete this form before submitting your online order for this item. Commercial customers or customers interested in using this enzyme for commercial purchases must purchase a license from Illumina prior to purchase.
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