HyperMu™ Transposase

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Last modified: 2008-07-15 14:57:15

HyperMu™ Transposase is a hyperactive enzyme that is at least 50 times more active than the MuA transposase available from other suppliers of Mu-based sytems. The highly purified, single-subunit HyperMu™ Transposase can be used to randomly insert (transpose or "hop") any HyperMu™ Transposon into any target DNA in vitro with an efficiency up to >10⁶ insertion clones per standard reaction. HyperMu™ Transposase can also catalyze transposition of other artificial Mu Transposons that have the R1 and R2 recognition sequences in the proper orientation at each end. The DNA between the R1 and R2 recognition sequences can be any DNA, including DNA encoding selectable markers, origins of replication, or other genetic elements.

Further, when HyperMu™ Transposase is incubated with an artificial Mu transposon in the absence of Mg²⁺, a stable HyperMu™ Transposome™ complex* is formed. Like an EZ-Tn5™ Transposome™ complex, a HyperMu™ Transposome can be electroporated into living cells, where it is activated by intracellular Mg²⁺ and the Mu transposon component is randomly inserted into the host's DNA in vivo.

A typical HyperMu transposition reaction requires four components:

1. HyperMu™ Transposase
2. Artificial Mu transposon
3. Target DNA
4. Presence of Mg²⁺

Insertion of an artificial Mu transposon into the target DNA results in a 5-bp duplication of target DNA sequence immediately adjacent to both ends of the transposon.

* 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 patent applications, exclusively licensed to Epicentre® Technologies.

Last modified: 2008-05-14 16:46:46

Applications

• In vitro insertion of an artificial Mu transposon into DNA cloned in vectors, such as plasmids, fosmids, cosmids, or BACs, as well as in vitro insertion into linear DNA.
• Preparation of HyperMu™ Transposomes for in vivo transposition of an artifical Mu transposon following electroporation into living cells.

Benefits

• Faster - completely sequence plasmid, cosmid, fosmid, and BAC clones 10-times faster than by subcloning or primer walking.
• Easier - a single reaction generates enough insertion clones to sequence even the largest BAC clone.
• More Economical - use only the sequencing primers provided in the kit to completely sequence your clone.
• Complete Sequence Coverage - insertion specificity is highly random to obtain the complete sequence of your DNA.

Last modified: 2008-05-14 16:46:46

Protocols for: HyperMu™ MuA Transposase

(catalogue number THM03210)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46