商品编号


TNP92110



品牌


Lucigen



公司


Lucigen



公司分类


In Vitro Tn5 Transposon Mutagenesis



Size

10 Units

商品信息


Create custom transposomes for
in vitro
transposomics

Integrate selectable
Marker
s, promoters or replication origins into DNA
in vitro
with EZ-Tn5 insertion kits





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.**

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

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
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.

Benefits

Specifically and uniquely recognizes 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 by
issued 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).

ORDER INFORMATION
10 units of 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.